Il-18 binding molecules

ABSTRACT

IL-18 participates in both innate and acquired immunity. The bioactivity of IL-18 is negatively regulated by the IL-18 binding protein (IL18BP), a naturally occurring and highly specific inhibitor. This soluble protein forms a complex with free IL-18 preventing its interaction with the IL-18 receptor, thus neutralizing and inhibiting its biological activity. The present invention discloses binding molecules, in particular antibodies or fragments thereof, which bind IL-18 and do not bind IL-18 bound to IL-18BP (IL-18/IL-18BP complex). Apart from its physiological role, IL-18 has been shown to mediate a variety of autoimmune and inflammatory diseases. The binding molecules of the inventions may be used as therapeutic molecules for treating IL-18-related autoimmune and inflammatory diseases or as diagnostic tools for characterizing, detecting and/or measuring IL-18 not bound to IL-18BP as component of the total IL-18 pool.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.16/111,659, filed Aug. 24, 2018 (U.S. Pat. No. 11,111,293), which is adivisional of U.S. patent application Ser. No. 15/164,508, filed May 25,2016 (U.S. Pat. No. 10,081,677), which is a divisional of U.S. patentapplication Ser. No. 14/017,561, filed Sep. 4, 2013 (U.S. Pat. No.9,376,489), which claims priority to U.S. Provisional Patent ApplicationNo. 61/697,981, filed Sep. 7, 2012, all of which are incorporated byreference in their entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Dec. 11, 2013 isnamed PAT055256-US-DIV03_SL.txt and is 226,528 bytes in size.

FIELD OF THE INVENTION

The present invention concerns binding molecules, more particularlyimmunoglobulins such as antibodies or fragments thereof, which bind withthe cytokine Interleukin 18 (IL-18) but do not bind IL-18 in complexwith the Interleukin 18 binding protein (IL-18BP) endogenous inhibitor.The present invention also concerns polynucleotide encoding said bindingmolecules, pharmaceutical compositions comprising said bindingmolecules, methods of treating and/or preventing diseases using saidbinding molecules and method for detecting and/or measuring the presenceand/or amount of IL-18 not bound to IL-18BP. Other aspects, objects andadvantages of the present invention will be apparent from thedescription below.

BACKGROUND OF THE INVENTION

Interleukin-18 (IL-18) was originally described in 1989 asinterferon-gamma inducing factor (IGIF). IL-18 is related to the IL-1family and is structurally related to IL-1β (Okamura H et al. (1995)Nature; 378:88-91). IL-18 is primarily produced by macrophages and Tcells as a precursor protein (pro-IL-18) and secreted as an activeprotein following cleavage by caspase-1 (Dinarello C A et al (1999) JAllergy Clin Immunol; 103:11-24). In normal physiology IL-18, in synergywith IL-12, is associated with induction of cell-mediated immunityfollowing infection with microbial products such as lipopolysaccharide(LPS) (Sareneva T et al (2000) J Immunol; 165(4):1933-8). Afterstimulation with IL-18, natural killer (NK) cells and T cells releasethe cytokine interferon gamma (INF-γ) which plays an important role inactivating macrophages and other cells. IL-18 has also various functionsin addition to an ability to induce interferon gamma. These biologicalproperties include activation of NF-κB, Fas ligand expression, theinduction of both CC and CXC chemokines, and increased production ofcompetent human immuno deficiency virus. Due to the ability of IL-18 toinduce INF-γ production in T cells and macrophages, it plays animportant role in Th1-type immune responses and participates in bothinnate and acquired immunity.

IL-18 binds with high affinity and signals through the IL-18 receptor(IL-18R), a heteromeric complex of alpha and beta chains encoded by thegenes IL18R1 and IL18RAP, respectively (Torigoe K et al (1997) J BiolChem; 272(41):25737-42). The bioactivity of IL-18 is negativelyregulated by the IL-18 binding protein (IL18BP), a naturally occurringand highly specific inhibitor. This soluble protein forms a complex withfree IL-18 preventing its interaction with the IL-18 receptor, thusneutralizing and inhibiting its biological activity (Dinarello C A(2000) Ann Rheum Dis; 59 Suppl 1:i17-20). IL-18BP is a constitutivelysecreted protein with high affinity binding to IL-18. Alternate mRNAsplicing variants of IL-18BP result in four isoforms. The prominent ‘a’isoform is present in the serum of healthy humans at 20-fold molarexcess compared with IL-18 (Dinarello and Kaplanski (2005) Expert RevClin Immunol, 1(4), 619-632). Apart from its physiological role, IL-18has been shown to mediate a variety of autoimmune and inflammatorydiseases. It has been demonstrated that IL-18 expression is up-regulatedin several autoimmune diseases, such as Crohn's disease, psoriasis,rheumatoid arthritis, multiple sclerosis and cardiovascular diseases(Braddock et al. (2004) Expert Opin Biol Ther; 4(6):847-860). IL-18 isalso up-regulated in certain inflammatory diseases such as chronicobstructive pulmonary disease (COPD) (Imaoka et al. (2008) Eur Respir;J31:287-297), idiopathic pulmonary fibrosis (IPF) (Kitasato et al.(2004) Am J Resp Cell Mol Biol; 31:619-625), macrophage activationsyndrome (MAS) (Dinarello and Kaplanski (2005) Expert Rev Clin Immunol;1(4): 619-632), adult onset Still's disease (AOSD) (Arlet J B et al.(2006) Ann Rheum Dis 65(12):1596-601) and systemic juvenile idiopathicarthritis (SJIA) (Akashi et al. (1994) Br J Haematol; 87(2):243-50).

Recent studies have shown that high amounts of IL-18 and INF-γ in bothinherited and acquired forms of hemophagocytic lymphohistiocytosis(HLH). HLH is characterized by activated lymphocytes and histiocytessecreting high amounts of inflammatory cytokines (Janka G E et al (2007)Eur J Paediatr; 166:95-109) and by impaired function of NK cells andcytotoxic T-cells. Most important, it has been shown that both forms arecharacterized by dis-regulation of IL-18 (Mazodier et al (2005)Immunobiology 106(10):3483-89).

Developing therapeutic molecules for targets such as IL-18 which areregulated by natural inhibitors can be very challenging. The presence ofan increased amount of systemic or local total IL-18 does not alwaysreflect the level of biologically active protein (IL-18 free ofIL-18BP). A therapeutic compound binding both free IL-18 and IL-18 incomplex to IL-18BP, although potentially capable of neutralizing theactivity of IL-18, would be required in higher dose than one which canselectively bind only biologically active free IL-18. Therapeuticcompounds which need to be administered in high dose may lead to morepronounced side effects or become immunogenic. High dosages alsotranslate into high production costs.

Therapeutic compounds competing for IL-18 when bound to IL18BP maydisturb the delicate balances of free/active IL-18 and IL-18BPbound/inactive IL-18 existing in patients stricken withdiseases/disorders characterized by IL-18 dis-regulation.

Finally, investigating diseases characterized by dis-regulation of IL-18free, in the absence of a diagnostic tool capable of detecting and/ormeasuring IL-18 free from IL-18BP as component of the total IL-18 wouldbe very difficult.

SUMMARY OF THE INVENTION

In one aspect the present invention therefore provides for a bindingmolecule that specifically binds IL-18, wherein the binding moleculedoes not bind the IL-18/IL-18 binding protein (IL-18 BP) complex andwherein the binding molecule is not IL-18BP.

In one embodiment of this aspect, the binding molecule binds to an IL-18epitope on IL-18 as defined with reference to SEQ ID NO:1, wherein theepitope comprises amino acids Arg140 and Glu152.

In another embodiment of this aspect, the epitope may further compriseany one or more of amino acids Gln92, Pro93, Gly95, Pro143, Glu157 orGlu177.

In yet another embodiment of this aspect, the epitope may furthercomprise any one or more of amino acids Lys89, Arg94, Met96, Phe138,Ser141, Gly144, His145, Asp146, Gln150 or Leu180.

In another embodiment of this aspect, the binding molecule does notcompete with IL-18BP for binding IL-18 when IL-18BP is bound to IL-18.

In another embodiment of this aspect the binding molecule is selectedfrom: an isolated antibody, a fragment of an isolated antibody, a singlevariable domain antibody, a bi- or multi-specific antibody, amultivalent antibody, a dual variable domain antibody, animmuno-conjugate, a fibronectin molecule, an adnectin, an DARPin, anavimer, an affibody, an anticalin, an affilin, a protein epitope mimeticor combinations thereof.

In another embodiment of this aspect, the binding molecule binds IL-18,wherein IL-18 comprises from amino acid 37 to amino acid 193 of SEQ IDNO:1 or SEQ ID NO:2.

In another embodiment of this aspect, the binding molecule inhibitsIL-18-dependent interferon gamma (INF-γ) production.

In another embodiment of this aspect, the binding molecule binds IL-18with a K_(D) of 100 pM or less.

In another embodiment of this aspect, the binding molecule according tothe invention is an isolated fully human, humanized or chimeric antibodyor a fragment thereof, preferably, an isolated fully human antibody.

In another embodiment of this aspect, the binding molecule is anantibody fragment or a single variable domain antibody, preferably aFab, a Fab′, a F(ab′)₂, a scFv, a dAb or a VHH.

In another embodiment of this aspect, the binding molecule is anisolated bispecific antibody or fragment thereof comprising a firstspecificity to IL-18 and a second specificity to another polypeptide,e.g. IL-12 or IL-1β.

In yet another embodiment of this aspect, the binding molecule is anisolated antibody comprising a mutated or chemically modified amino acidFc region, wherein the mutated or chemically modified amino acid Fcregion prevents or decreases ADCC activity and/or increase half lifewhen compared with a wild type Fc region. Preferably, the mutated orchemically modified amino acid Fc region is a silent IgG1 Fc region.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO: 3        or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO: 4        or SEQ ID NO: 9 or SEQ ID NO: 10 or SEQ ID NO: 11 or SEQ ID NO:        12 or SEQ ID NO: 13 or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        5 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO: 6        or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO: 7        or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO: 8        or conservative variants thereof.

Preferably, the antibody or fragment thereof comprises a heavy chainvariable region H-CDR 2 comprising SEQ ID NO: 9 or SEQ ID NO: 13.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises a light chain variable domaincomprising SEQ ID NO: 16 or SEQ ID NO: 20 or conservative variantsthereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable domain comprising SEQ ID NO: 14 or SEQ        ID NO: 22 or SEQ ID NO: 25 or SEQ ID NO: 28 or SEQ ID NO: 31 or        SEQ ID NO: 34 or conservative variants thereof and a light chain        variable domain comprising SEQ ID NO: 16 or conservative        variants thereof or    -   ii. a heavy chain variable domain comprising SEQ ID NO: 18 or        SEQ ID NO: 37 or SEQ ID NO: 40 or conservative variants thereof        and a light chain variable domain comprising SEQ ID NO: 20 or        conservative variants thereof.

Preferably, the heavy chain variable domain comprises SEQ ID NO: 14 orconservative variants thereof and a light chain variable domaincomprises SEQ ID NO: 16 or conservative variants thereof and,optionally, amino acid lysine (Lys; K) in position 30 with reference toSEQ ID NO:14 is replaced by an amino acid selected from asparagine (Asn;N) or serine (Ser; S) or threonine (Thr; T) or alanine (Ala; A) orglutamate (Glu; E) or histidine (His; H) or leucine (Leu; L) orglutamine (Gln; Q) or arginine (Arg; R) or valine (Val; V) or tyrosine(Tyr; Y) or isoleucine (Ile; I).

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises a heavy chain variable domaincomprising SEQ ID NO: 18 or conservative variants thereof and a lightchain variable domain comprising SEQ ID NO: 20 or conservative variantsthereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex, the isolated antibody or a fragmentthereof comprises:

-   -   i. a heavy chain comprising SEQ ID NO: 43 or SEQ ID NO: 47 or        SEQ ID NO: 50 or SEQ ID NO: 56 or conservative variants thereof        and a light chain comprising SEQ ID NO: 45 or conservative        variants thereof or    -   ii. a heavy chain comprising SEQ ID NO: 53 or SEQ ID NO: 100 or        SEQ ID NO: 158 or conservative variants thereof and a light        chain comprising SEQ ID NO: 160 or conservative variants        thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex, the isolated antibody or a fragmentthereof comprises:

-   -   i. a heavy chain comprising SEQ ID NO: 43 or conservative        variants thereof and a light chain comprising SEQ ID NO: 45 or        conservative variants thereof or    -   ii. a heavy chain comprising SEQ ID NO: 158 or conservative        variants thereof and a light chain comprising SEQ ID NO: 160 or        conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO: 74        or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO:        75 or SEQ ID NO: 76 or SEQ ID NO: 77 or SEQ ID NO: 78 or        conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        79 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO:        80 or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO: 81        or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO:        82 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises a light chain variable domaincomprising SEQ ID NO: 85 or conservative variants thereof and a heavychain variable domain comprising SEQ ID NO: 83 or SEQ ID NO: 87 or SEQID NO: 90 or SEQ ID NO: 93 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex, the isolated antibody comprises aheavy chain comprising SEQ ID NO: 96 or SEQ ID NO: 103 or conservativevariants thereof and a light chain comprising SEQ ID NO: 98 orconservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO:        106 or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO:        107 or SEQ ID NO: 122 or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        108 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO:        109 or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO:        110 or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO:        111 or SEQ ID NO: 126 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO:        106 conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO:        107 or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        108 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO:        109 or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO:        110 or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO:        111 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable domain comprising SEQ ID NO: 112 or        conservative variants thereof and a light chain variable domain        comprising SEQ ID NO: 114 or conservative variants thereof or    -   ii. a heavy chain variable domain comprising SEQ ID NO: 138 or        conservative variants thereof and a light chain variable domain        comprising SEQ ID NO: 140 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain comprising SEQ ID NO: 116 or conservative        variants thereof and a light chain comprising SEQ ID NO: 118 or        conservative variants thereof or    -   ii. a heavy chain comprising SEQ ID NO: 142 or conservative        variants thereof and a light chain comprising SEQ ID NO: 144 or        conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO:        120 or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO:        121 or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        123 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO:        124 or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO:        125 or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO:        127 or SEQ ID NO: 128 or SEQ ID NO: 129 or conservative variants        thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex, the isolatedantibody or a fragment thereof comprises a heavy chain variable domaincomprising SEQ ID NO: 130 or conservative variants thereof and a lightchain variable domain comprising SEQ ID NO: 132 or SEQ ID NO: 147 or SEQID NO: 153 or conservative variants thereof.

In another embodiment, when the binding molecule of the invention is anisolated antibody which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex, the isolated antibody comprises aheavy chain comprising SEQ ID NO: 134 or conservative variants thereofand a light chain comprising SEQ ID NO: 136 or SEQ ID NO: 150 or SEQ IDNO: 156 or conservative variants thereof.

In another aspect, there is provided an isolated polynucleotide encodingthe binding molecule according to the invention.

In one embodiment of this later aspect, the isolated polynucleotideaccording to the invention encodes a heavy chain variable domain,wherein the polynucleotide:

-   -   i. is at least 90% identical to SEQ ID NO: 15 or SEQ ID NO: 19        or SEQ ID NO: 23 or SEQ ID NO: 26 or SEQ ID NO: 29 or SEQ ID NO:        32 or SEQ ID NO: 35 or SEQ ID NO: 38 or SEQ ID NO: 41 or SEQ ID        NO: 84 or SEQ ID NO: 88 or SEQ ID NO: 91 or SEQ ID NO: 94 or SEQ        ID NO: 113 or SEQ ID NO: 131 or SEQ ID NO: 139 or SEQ ID NO: 146        or SEQ ID NO: 152; or    -   ii. comprises SEQ ID NO: 15 or SEQ ID NO: 19 or SEQ ID NO: 23 or        SEQ ID NO: 26 or SEQ ID NO: 29 or SEQ ID NO: 32 or SEQ ID NO: 35        or SEQ ID NO: 38 or SEQ ID NO: 41 or SEQ ID NO: 84 or SEQ ID NO:        88 or SEQ ID NO: 91 or SEQ ID NO: 94 or SEQ ID NO: 113 or SEQ ID        NO: 131 or SEQ ID NO: 139 or SEQ ID NO: 146 or SEQ ID NO: 152;        or    -   iii. consists essentially of SEQ ID NO: 15 or SEQ ID NO: 19 or        SEQ ID NO: 23 or SEQ ID NO: 26 or SEQ ID NO: 29 or SEQ ID NO: 32        or SEQ ID NO: 35 or SEQ ID NO: 38 or SEQ ID NO: 41 or SEQ ID NO:        84 or SEQ ID NO: 88 or SEQ ID NO: 91 or SEQ ID NO: 94 or SEQ ID        NO: 113 or SEQ ID NO: 131 or SEQ ID NO: 139 or SEQ ID NO: 146 or        SEQ ID NO: 152.

In another embodiment of this aspect, the isolated polynucleotideaccording to the invention encodes a light chain variable domain,wherein the polynucleotide:

-   -   i. is at least 90% identical to SEQ ID NO: 17 or SEQ ID NO: 21        or SEQ ID NO: 24 or SEQ ID NO: 27 or SEQ ID NO: 30 or SEQ ID NO:        33 or SEQ ID NO: 36 or SEQ ID NO: 39 or SEQ ID NO: 42 or SEQ ID        NO: 86 or SEQ ID NO: 89 or SEQ ID NO: 92 or SEQ ID NO: 95 or SEQ        ID NO: 115 or SEQ ID NO: 133 or SEQ ID NO: 141 or SEQ ID NO: 148        or SEQ ID NO: 154; or    -   ii. comprises SEQ ID NO: 17 or SEQ ID NO: 21 or SEQ ID NO: 24 or        SEQ ID NO: 27 or SEQ ID NO: 30 or SEQ ID NO: 33 or SEQ ID NO: 36        or SEQ ID NO: 39 or SEQ ID NO: 42 or SEQ ID NO: 86 or SEQ ID NO:        89 or SEQ ID NO: 92 or SEQ ID NO: 95 or SEQ ID NO: 115 or SEQ ID        NO: 133 or SEQ ID NO: 141 or SEQ ID NO: 148 or SEQ ID NO: 154;        or    -   iii. consists essentially of SEQ ID NO: 17 or SEQ ID NO: 21 or        SEQ ID NO: 24 or SEQ ID NO: 27 or SEQ ID NO: 30 or SEQ ID NO: 33        or SEQ ID NO: 36 or SEQ ID NO: 39 or SEQ ID NO: 42 or SEQ ID NO:        86 or SEQ ID NO: 89 or SEQ ID NO: 92 or SEQ ID NO: 95 or SEQ ID        NO: 115 or SEQ ID NO: 133 or SEQ ID NO: 141 or SEQ ID NO: 148 or        SEQ ID NO: 154.

In another embodiment of this aspect, the isolated polynucleotideaccording to the invention encodes a heavy chain, wherein thepolynucleotide:

-   -   i. is at least 90% identical to SEQ ID NO: 44 or SEQ ID NO: 48        or SEQ ID NO: 51 or SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO:        97 or SEQ ID NO: 101 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ        ID NO: 135 or SEQ ID NO: 143 or SEQ ID NO: 149 or SEQ ID NO: 155        or SEQ ID NO: 159; or    -   ii. comprises SEQ ID NO: 44 or SEQ ID NO: 48 or SEQ ID NO: 51 or        SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO: 97 or SEQ ID NO:        101 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID NO: 135 or SEQ        ID NO: 143 or SEQ ID NO: 149 or SEQ ID NO: 155 or SEQ ID NO:        159; or    -   iii. consists essentially of SEQ ID NO: 44 or SEQ ID NO: 48 or        SEQ ID NO: 51 or SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO: 97        or SEQ ID NO: 101 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID        NO: 135 or SEQ ID NO: 143 or SEQ ID NO: 149 or SEQ ID NO: 155 or        SEQ ID NO: 159.

In another embodiment of this aspect, the isolated polynucleotideaccording to the invention encodes a light chain, wherein thepolynucleotide:

-   -   i. is at least 90% identical to SEQ ID NO: 46 or SEQ ID NO: 49        or SEQ ID NO: 52 or SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO:        99 or SEQ ID NO: 102 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ        ID NO: 137 or SEQ ID NO: 145 or SEQ ID NO: 151 or SEQ ID NO: 157        or SEQ ID NO: 161; or    -   ii. comprises SEQ ID NO: 46 or SEQ ID NO: 49 or SEQ ID NO: 52 or        SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO: 99 or SEQ ID NO:        102 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ ID NO: 137 or SEQ        ID NO: 145 or SEQ ID NO: 151 or SEQ ID NO: 157 or SEQ ID NO:        161; or    -   iii. consists essentially of SEQ ID NO: 46 or SEQ ID NO: 49 or        SEQ ID NO: 52 or SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO: 99        or SEQ ID NO: 102 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ ID        NO: 137 or SEQ ID NO: 145 or SEQ ID NO: 151 or SEQ ID NO: 157 or        SEQ ID NO: 161.

In another aspect of this aspect, it is provided a cloning or expressionvector comprising one or more polynucleotides as claimed herein.

In one embodiment, the cloning or expression vector according to theinvention comprises at least one polynucleotide selected from the groupof SEQ ID NO: 44 or SEQ ID NO: 48 or SEQ ID NO: 51 or SEQ ID NO: 54 orSEQ ID NO: 57 or SEQ ID NO: 101 or SEQ ID NO: 159 or SEQ ID NO: 97 orSEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID NO: 143 or SEQ ID NO: 135 orSEQ ID NO: 149 or SEQ ID NO: 155 or SEQ ID NO: 46 or SEQ ID NO: 49 orSEQ ID NO: 52 or SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO: 102 or SEQID NO: 161 or SEQ ID NO: 99 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQID NO: 145 or SEQ ID NO: 137 or SEQ ID NO: 151 or SEQ ID NO: 157.

The present invention further provides for a host cell comprising one ormore cloning or expression vectors as claimed herein.

In another aspect of the present invention there is provided a stablytransformed or transfected host cell comprising one or morepolynucleotides as claimed herein.

The present invention further provides for a method of producing abinding molecule, which method comprises culturing a host cell asclaimed herein under conditions suitable for producing the bindingmolecule.

In another aspect the present invention further provides for apharmaceutical composition comprising a pharmaceutical carrier and thebinding molecule or the isolated antibody or a fragment thereof whichbinds IL-18 and does not bind the IL-18/IL-18 binding protein (IL-18 BP)complex and wherein the binding molecule is not IL-18BP.

Preferably, the pharmaceutical composition according to the invention isin intravenously, inhalable or sub-cutaneously administrable form.

The present invention further provide for a binding molecule which bindsIL-18 and does not bind the IL-18/IL-18 binding protein (IL-18 BP)complex and wherein the binding molecule is not IL-18BP or thepharmaceutical composition comprising that binding molecule for use intherapy.

In another aspect of the present invention there is provided a bindingmolecule which binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and wherein the binding molecule is notIL-18BP or the pharmaceutical composition comprising that bindingmolecule for use in treating and/or preventing sarcoidosis, inparticular pulmonary sarcoidosis, hemophagocytic lymphohistiocytosis(HLH), familial hemophagocytic lymphohistiocytosis (FHL) and otherimmunodeficiency syndromes, Giant Cell Arthritis (GCA), chronicobstructive pulmonary disease (COPD), adult onset Still's Disease(AOSD), systemic juvenile idiopathic arthritis (SJIA), severe asthma,Uveitis, Geographic Atrophy, diabetes type 1, diabetes type 2 oratherosclerosis and any combination thereof in a mammalian patient.

In another aspect of the present invention there is provided a method oftreating and/or preventing sarcoidosis, in particular pulmonarysarcoidosis, hemophagocytic lymphohistiocytosis (HLH), familialhemophagocytic lymphohistiocytosis (FHL) and other immunodeficiencysyndromes, Giant Cell Arthritis (GCA), chronic obstructive pulmonarydisease (COPD), adult onset Still's Disease (AOSD), systemic juvenileidiopathic arthritis (SJIA), severe asthma, Uveitis, Geographic Atrophy,diabetes type 1, diabetes type 2 or atherosclerosis and any combinationthereof in a mammalian patient which method comprises administrating tothe mammalian patient a therapeutically effective amount of a bindingmolecule which binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and wherein the binding molecule is notIL-18BP or the pharmaceutical composition comprising that bindingmolecule.

In yet another aspect of the present invention there is provided the useof the binding molecule which binds IL-18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex and wherein the bindingmolecule is not IL-18BP or the pharmaceutical composition comprisingthat binding molecule in the manufacture of a medicament for treatingand/or preventing sarcoidosis, in particular pulmonary sarcoidosis,hemophagocytic lymphohistiocytosis (HLH), familial hemophagocyticlymphohistiocytosis (FHL) and other immunodeficiency syndromes, GiantCell Arthritis (GCA), chronic obstructive pulmonary disease (COPD),adult onset Still's Disease (AOSD), systemic juvenile idiopathicarthritis (SJIA), severe asthma, Uveitis, Geographic Atrophy, diabetestype 1, diabetes type 2 or atherosclerosis and any combination thereofin a mammalian patient.

In one embodiment according to the use of the present invention, themammalian patient is a human patient.

In another aspect, there is provided for a complex comprising IL-18 anda binding molecule which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and wherein the binding molecule isnot IL-18BP according to the invention.

In another aspect, the invention provides for a binding molecule orisolated antibody or a fragment thereof which binds IL-18 and does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex and wherein thebinding molecule is not IL-18BP for use in diagnosis or for use in adiagnostic kit.

In yet another aspect, the present invention provides for a bindingmolecule or isolated antibody or a fragment thereof which binds IL-18and does not bind the IL-18/IL-18 binding protein (IL-18 BP) complex andwherein the binding molecule is not IL-18BP for use in diagnosis or foruse in detecting and/or measuring the presence and/or the amount of freeIL-18 (i.e. IL-18 not bound to IL-18BP) in a sample.

In yet a further aspect of the present invention, there is provided amethod for detecting and/or measuring the presence and/or amount of freeIL-18 (i.e. IL-18 not bound to IL-18BP) in a sample, wherein the sampleis optionally a human sample, wherein the method comprises contactingthe sample with the binding molecule or the isolated antibody or afragment thereof which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and wherein the binding molecule isnot IL-18BP.

In one embodiment of this aspect, the sample is human blood.

In another aspect, the present invention further provides for adiagnostic kit comprising the binding molecule or isolated antibody or afragment thereof which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and wherein the binding molecule isnot IL-18BP and/or the complex comprising IL-18 and that bindingmolecule wherein the kit optionally comprises a first control compound.

In one embodiment of this aspect, the first control compound is freeIL-18 and the kit optionally comprises a second control compound whichis murine antibody 125-2H.

In another aspect of the present invention, there is provided a medicalor diagnostic device comprising the binding molecule or the isolatedantibody or a fragment thereof which binds IL-18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex and wherein the bindingmolecule is not IL-18BP and/or the complex comprising IL-18 and thatbinding molecule.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: CDRs of the heavy chains variable regions of the antibodiesexemplified herein.

FIG. 2: CDRs of the light chains variable regions of the antibodiesexemplified herein.

FIG. 3 (A-D): Effect of the anti-IL-18 antibodies and fragments thereofof the present invention on IFN-γ release from PBMCs induced byLPS/IL-12 (stimulation by native IL-18). Data showsconcentration-dependent inhibition of native IL-18-induced IFN-γ releasefrom freshly isolated human PBMCs from individual donors, where eachdata point represents mean±SEM from n=4 wells. Native IL-18 wasstimulated by LPS/IL-12 treatment and the dotted line represents theextent of IL-18-dependency, as determined by maximal efficacy of humanIL-18BPa-Fc.

FIG. 4 (A-B): Effect of anti-IL-18 antibodies and fragments thereof ofthe present invention on IFN-γ release from PBMCs induced by recombinantIL-18. Data shows concentration-dependent inhibition of recombinanthuman IL-18 (1 nM)-induced IFN-γ release from freshly isolated humanPBMCs from representative individual donors, where each data pointrepresents mean±SEM from n=4 wells.

FIG. 5 (A-C): Effect of anti-IL-18 antibodies and fragments thereof ofthe present invention on IFN-γ release from KG-1 cells induced by humanIL-18. Data shows concentration-dependent inhibition of recombinanthuman IL-18 (1 nM)-induced IFN-γ release from KG-1 cells, where eachdata point represents mean±SEM from n=4 wells.

FIG. 6 (A-C): Effect of anti-IL-18 antibodies and fragments thereof ofthe present invention on IFN-γ release from KG-1 cells induced bycynomolgus IL-18. Data shows concentration-dependent inhibition ofrecombinant cynomolgus IL-18 (0.2 nM)-induced IFN-γ release from KG-1cells, where each data point represents mean±SEM from n=4 wells.

FIG. 7 (A-E): Effect of anti-IL-18 antibodies and fragments thereof ofthe present invention on human IL-18-induced by LPS/IL-12 (stimulationof native IL-18) and subsequent IFN-γ release in human whole blood.

FIG. 8 (A-E): Effect of anti-IL-18 antibodies and fragments thereof ofthe present invention on human IL-18-induced IFN-γ release in humanwhole blood. Data shows concentration-dependent inhibition ofrecombinant human IL-18-induced IFN-γ release from whole blood takenfrom individual donors, where each data point represents mean±SEM fromn=4 wells.

FIG. 9 (A-B): Binding of anti-IL-18 antibodies and fragments toIL-18/IL-18BP complex. (A) Parental MOR08775 and MOR08776 do notrecognize the IL-18/IL-18BP complex. In this experiment, MOR08775 andMOR08776 were incubated with biotinylated IL-18/IL-18BP complex. (B) Theexperiment was performed in a similar way as shown in A), except forusing unbiotinylated human IL-18. MOR03207 is an anti-lysozyme antibodyand 125-2H is anti-IL-18 mouse IgG which binds the IL-18/IL-18BPcomplex.

FIG. 10: Epitope binning for anti-IL-18 antibodies MOR8775, MOR8776 (A))and for antibodies MOR9464, MOR9464_N30K, MOR10222_N30S_M54I andMOR14431 (B) Black cell fillings indicate antibodies competing for asimilar epitope, whilst no cell filling indicates no competition.

FIG. 11 (A-B): A) Structure of the IL-1β/IL-1R complex superposed ontothe model of the IL-18/IL-18Rα created on the structure of theIL-1β/IL-1R complex. All the structures have been represented as ribbon.The structures of IL-18 and IL-1β have been used for thesuperimposition. B) Model of the IL-18/IL-18Rα complex created on thestructure of the IL-1β/IL-1R complex. IL-18Rα (shown in surfacerepresentation) comprises three immunoglobulin-like domains, D1, D2 andD3. On IL-18 (shown in ribbon representation) sites 1 and 2 are theIL-18Rα binding sites. Site 3 is the IL-18Rβ binding site.

FIG. 12: Comparisons of the relevant amino acids bound on IL-18. 1)Sequence of IL-18 with reference to SEQ ID NO:1 from amino acids 37 to193. 2) from Kim et al., (2000) Proc Natl Acad Sci; 97(3):1190-1195(modelling of IL-18BP and its complex with human IL-18). 3) from Krummet al., (2008) Proc Natl Acad Sci; 2008 105(52): 20711-2071552, Table 1.4) from Kato et al., (2003) Nature Struct. Biol. 2003; 10(11): 966-971;residues involved in IL-18Rα binding. 5) H/DxMS results for MOR9464bound to IL-18.

FIG. 13: Overall view of the three-dimensional structure of the complexbetween human IL-18 (shown in Cα trace and solvent accessible surface)and MOR9464_N30K (shown in cartoon representation).

FIG. 14 (A-B): (A) sequence alignment of the sequences of mature humanand cynomolgus IL-18 (from amino acid 37 to 193); (B) Space-fillingrepresentation of the 6 amino acids which differ across the two species.E177 is the only one present in the antibody complex interface. IL-18 isshown in Cα trace and solvent accessible surface and MOR9464_N30K isshown in cartoon representation.

FIG. 15: Ribbon representation of the IL18/MOR9464_N30K complexsuperposed onto the model of the IL-18/IL-18Rα complex created on thestructure of the IL-1β/IL-1R complex. IL-18Rα structure is shown with asurface representation and the MOR9464_N30K heavy and light chains arein dark and light grey respectively.

FIG. 16: Ribbon representation of human IL-18 bound to IL-18 BP frompoxvirus (left) and to MOR9464_N30K antibody fragment (right) and theirsuperposition (centre). Overlay is based on the IL-18 structure. IL-18is shown in Cα trace and solvent accessible surface and MOR9464_N30K andIL-18 BP from poxvirus are shown in cartoon representation.

FIG. 17: Ribbon representation of human IL-18 bound to MOR9464_N30Kantibody fragment (right) and to murine 125-2H antibody fragment (right)and their superposition (centre). Overlay is based on the IL-18structure. IL-18 is shown in Cα trace and solvent accessible surface andMOR9464_N30K and 125-2H are shown in cartoon representation

FIG. 18: Ribbon representation of human IL-18 bound to 125-2H overlaidonto IL-18 bound to IL-18BP from poxvirus. Overlay is based on the IL-18structure. IL-18 is shown in Cα trace and solvent accessible surface andMOR9464_N30K, 125-2H and IL-18BP from poxvirus are shown in cartoonrepresentation.

FIG. 19: Ribbon representation of human IL-18 bound to MOR9464_N30Kantibody fragment, with epitope and paratope residues depicted in stickrepresentation, showing the specific interaction with amino acid lysineat position 30 of MOR9464_N30K.

DETAILED DESCRIPTION OF THE INVENTION 1. Definitions

For purposes of interpreting this specification, the followingdefinitions will apply and whenever appropriate, terms used in thesingular will also include the plural and vice versa. Additionaldefinitions are set forth throughout the detailed description.

The term “IL-18” is synonym to IL-18 polypeptide, Interleukin-18polypeptide, IFN-gamma-inducing factor orInterferon-gamma-inducing-factor or INF-γ inducing factor. The term“IL-18” refers to human IL-18 comprising amino acids 37 to 193 of SEQ IDNO: 1. Throughout this specification, the term IL-18 encompasses bothpro-IL-18 (precursor of mature IL-18 prior protease cleavage) and matureIL-18 (post protease cleavage) interchangeably unless it is specifiedthat the pro- or mature form is meant.

The term cm IL-18 refers to cynomolgus monkey IL-18 comprising aminoacids 37 to 193 of SEQ ID NO:2.

The term “antibody” refers to an intact immunoglobulin or a functionalfragment thereof. Naturally occurring antibodies typically comprise atetramer which is usually composed of at least two heavy (H) chains andat least two light (L) chains. Each heavy chain is comprised of a heavychain variable region (abbreviated herein as VH) and a heavy chainconstant region, usually comprised of three domains (CH1, CH2 ad CH3).Heavy chains can be of any isotype, including IgG (IgG1, IgG2, IgG3 andIgG4 subtypes), IgA (IgA1 and IgA2 subtypes), IgM and IgE. Each lightchain is comprised of a light chain variable region (abbreviated hereinas VL) and a light chain constant region (CL). Light chain includeskappa chains and lambda chains. The heavy and light chain variableregion is typically responsible for antigen recognition, whilst theheavy and light chain constant region may mediate the binding of theimmunoglobulin to host tissues or factors, including various cells ofthe immune system (e.g., effector cells) and the first component (Clq)of the classical complement system. The VH and VL regions can be furthersubdivided into regions of hypervariability, termed complementaritydetermining regions (CDR), interspersed with regions that are moreconserved, termed framework regions (FR). Each VH and VL is composed ofthree CDRs and four FRs arranged from amino-terminus to carboxy-terminusin the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Thevariable regions of the heavy and light chains contain a binding domainthat interacts with an antigen.

The term “antigen-binding portion” of an antibody (or simply “antigenportion”), as used herein, refers to full length or one or morefragments of an antibody that retain the ability to specifically bind toIL-18. It has been shown that the antigen-binding function of anantibody can be performed by fragments of a full-length antibody.Examples of binding fragments encompassed within the term“antigen-binding portion” of an antibody include a Fab fragment, amonovalent fragment consisting of the VL, VH, CL and CH1 domains; aF(ab)2 fragment, a bivalent fragment comprising two Fab fragments linkedby a disulfide bridge at the hinge region; a Fd fragment consisting ofthe VH and CH1 domains; a Fv fragment consisting of the VL and VHdomains of a single arm of an antibody; a dAb fragment (Ward et al.,(1989) Nature; 341:544 546), which consists of a VH domain; and anisolated complementarity determining region (CDR).

Furthermore, although the two domains of the Fv fragment, VL and VH, arecoded for by separate genes, they can be joined, using recombinantmethods, by a flexible linker that enables them to be made as a singleprotein chain in which the VL and VH regions pair to form monovalentmolecules (known as single chain Fv (scFv); see e.g., Bird et al.,(1988) Science 242:423-426; and Huston et al., (1988) Proc Natl Acad Sc;85:5879-5883). Such single chain antibodies are also intended to beencompassed within the term “antigen-binding portion” of an antibody.These antibody fragments are obtained using conventional techniquesknown to those of skill in the art, and the fragments are screened forutility in the same manner as are intact antibodies.

The term “isolated” means throughout this specification, that theimmunoglobulin, antibody or polynucleotide, as the case may be, existsin a physical milieu distinct from that in which it may occur in nature.

An isolated antibody that specifically binds the IL18 polypeptide may,however, have cross-reactivity to other antigens, such as IL18 fromother species (e.g. cynomolgus monkey (cm) IL-18). Moreover, an isolatedantibody may be substantially free of other cellular material and/orchemicals.

Throughout this specification, complementarity determining regions(“CDR”) are defined according to the Kabat definition unless specifiedthat the CDR are defined according to the Chothia definition or by bothdefinitions together. The Kabat definition is a standard for numberingthe residues in an antibody and it is typically used to identify CDRregions (Kabat et al., (1991), 5th edition, NIH publication No.91-3242). The Chothia definition is similar to the Kabat definition butit takes into account positions of certain structural loops (Chothia etal., (1987) J. Mol. Biol., 196:901-17; Al-Lazikani et al., (1997) J.Mol. Biol. 273:927-948). By convention, the CDR regions in the heavychain are typically referred to as H-CDR1, H-CDR2 and H-CDR3 and in thelight chain as L-CDR1, LCDR2 and L-CDR3. They are numbered sequentiallyin the direction from the amino terminus to the carboxy terminus.

The terms “monoclonal antibody” or “monoclonal antibody composition” asused herein refer to a preparation of antibody molecules of singlemolecular composition. A monoclonal antibody composition displays asingle binding specificity and affinity for a particular epitope.

The term “human antibody”, as used herein, is intended to includeantibodies having variable regions in which both the framework and CDRregions are derived from sequences of human origin. Furthermore, if theantibody contains a constant region, the constant region also is derivedfrom such human sequences, e.g., human germline sequences, or mutatedversions of human germline sequences or antibody containing consensusframework sequences derived from human framework sequences analysis, forexample, as described in Knappik, et al., (2000) J Mol Biol; 296:57-86).

The human antibodies of the invention may include amino acid residuesnot encoded by human sequences (e.g., mutations introduced by random orsite-specific mutagenesis in vitro or by somatic mutation in vivo).However, the term “human antibody”, as used herein, is not intended toinclude antibodies in which CDR sequences derived from the germline ofanother mammalian species, such as a mouse, have been grafted onto humanframework sequences.

The term “human monoclonal antibody” refers to antibodies displaying asingle binding specificity which have variable regions in which both theframework and CDR regions are derived from human sequences.

The term “recombinant human antibody”, as used herein, includes allhuman antibodies that are prepared, expressed, created or isolated byrecombinant means, such as antibodies isolated from an animal (e.g., amouse) that is transgenic or transchromosomal for human immunoglobulingenes or a hybridoma prepared therefrom, antibodies isolated from a hostcell transformed to express the human antibody, e.g., from atransfectoma, antibodies isolated from a recombinant, combinatorialhuman antibody library, and antibodies prepared, expressed, created orisolated by any other means that involve splicing of all or a portion ofa human immunoglobulin gene. Such recombinant human antibodies havevariable regions in which the framework and CDR regions are derived fromhuman germline immunoglobulin sequences. In certain embodiments,however, such recombinant human antibodies can be subjected to in vitromutagenesis (or, when an animal transgenic for human Ig sequences isused, in vivo somatic mutagenesis) and thus the amino acid sequences ofthe VH and VL regions of the recombinant antibodies are sequences that,while derived from and related to human germline VH and VL sequences,may not naturally exist within the human antibody germline repertoire invivo.

The phrases “an antibody recognizing an antigen” and “an antibodyspecific for an antigen” are used interchangeably herein with the term“an antibody which binds specifically to an antigen”.

As used herein, a binding molecule that “specifically binds to IL-18” isintended to refer to a binding molecule that binds to human IL-18 with aK_(D) of a 100 nM or less, 10 nM or less, 1 nM or less.

A binding molecule that “cross-reacts with an antigen other than IL-18is intended to refer to a binding molecule that binds that antigen witha K_(D) of a 100 nM or less, 10 nM or less, 1 nM or less. A bindingmolecule that “does not cross-react with a particular antigen” isintended to refer to a binding molecule that exhibits essentiallyundetectable binding against these proteins in standard binding assays.

As used herein, the term “antagonist” is intended to refer to a bindingmolecule that inhibits IL-18 dependent signalling activity in thepresence of IL-18 in a human cell assay such as IL-18 dependentInterferon-gamma (IFN-γ) production assay in human blood cells. Examplesof an IL-18 dependent IFN-γ production assay in human blood cells aredescribed in more details in the examples below.

As used herein, an antibody with “no agonistic activity” is intended torefer to a binding molecule that does not significantly increase IL-18dependent signalling activity in the absence and/or presence of IL-18 ina cell-based assay, such as human blood cells IFN-γ production assay.Such assays are described in more details in the examples below.

The term “K_(assoc)” or “K_(a)”, as used herein, is intended to refer tothe association rate of a particular binding molecule-antigeninteraction, whereas the term “K_(dis)” or “K_(d),” as used herein, isintended to refer to the dissociation rate of a particular bindingmolecule-antigen interaction. The term “K_(D)”, as used herein, isintended to refer to the dissociation constant, which is obtained fromthe ratio of K_(d) to K_(a) (i.e. K_(d)/K_(a)) and is expressed as amolar concentration (M). K_(D) values for antibodies can be determinedusing methods well established in the art. A method for determining theK_(D) of an antibody is by using surface plasmon resonance, such as aBiacore® system.

As used herein, the term “affinity” refers to the strength ofinteraction between binding molecule and antigen at single antigenicsites.

As used herein, the term “high affinity” for an antibody refers to anantibody having a K_(D) of 1 nM or less for a target antigen.

As used herein, the term “subject” includes any human or non-humananimal.

The term “non-human animal” includes all vertebrates, e.g., mammals andnon-mammals, such as non-human primates, sheep, dogs, cats, horses,cows, chickens, amphibians, reptiles, etc.

As used herein, the term, “optimized nucleotide sequence” means that thenucleotide sequence has been altered to encode an amino acid sequenceusing codons that are preferred in the production cell or organism,generally a eukaryotic cell, for example, a cell of Pichia pastoris, aChinese Hamster Ovary cell (CHO) or a human cell. The optimizednucleotide sequence is engineered to retain completely the amino acidsequence originally encoded by the starting nucleotide sequence, whichis also known as the “parental” sequence. The optimized sequences hereinhave been engineered to have codons that are preferred in CHO mammaliancells; however optimized expression of these sequences in othereukaryotic cells is also envisioned herein.

The term “identity” refers to the similarity between at least twodifferent sequences. This identity can be expressed as a percentidentity and determined by standard alignment algorithms, for example,the Basic Local Alignment Tool (BLAST) (Altshul et al., (1990) J MolBiol; 215:403 410); the algorithm of Needleman et al., (1970) J MolBiol; 48:444-453 or the algorithm of Meyers et al., (1988) Comput ApplBiosci; 4:11-17). A set of parameters may be the Blosum 62 scoringmatrix with a gap penalty of 12, a gap extend penalty of 4, and aframeshift gap penalty of 5. The percent identity between two amino acidor nucleotide sequences can also be determined using the algorithm of E.Meyers and W. Miller, (1989) CABIOS; 4(1):1-17) which has beenincorporated into the ALIGN program (version 2.0), using a PAM 120weight residue table, a gap length penalty of 12 and a gap penalty of 4.The percent identity is usually calculated by comparing sequences ofsimilar length.

The term “immune response” refers to the action of, for example,lymphocytes, antigen presenting cells, phagocytic cells, granulocytes,and soluble macromolecules produced by the above cells or the liver(including antibodies, cytokines, and complement) that results inselective damage to, destruction of, or elimination from the human bodyof invading pathogens, cells or tissues infected with pathogens,cancerous cells, or, in cases of autoimmunity or pathologicalinflammation, normal human cells or tissues.

A “signal transduction pathway” or “signaling activity” refers to abiochemical causal relationship generally initiated by a protein-proteininteraction such as binding of a growth factor to a receptor, resultingin transmission of a signal from one portion of a cell to anotherportion of a cell. In general, the transmission involves specificphosphorylation of one or more tyrosine, serine, or threonine residueson one or more proteins in the series of reactions causing signaltransduction. Penultimate processes typically include nuclear events,resulting in a change in gene expression.

The term “neutralises” and grammatical variations thereof meansthroughout this specification, that the biological activity of thetarget is reduced either totally or partially in the presence of thebinding protein or antibody, as the case may be.

The term “nucleic acid” or “polynucleotide” refers to deoxyribonucleicacid (DNA) or ribonucleic acid (RNA) and polymers thereof in eithersingle- or double-stranded form. Unless specifically limited, the termencompasses nucleic acids containing known analogues of naturalnucleotides that have similar binding properties as the referencenucleic acid and are metabolized in a manner similar to naturallyoccurring nucleotides. Unless otherwise indicated, a particular nucleicacid sequence also implicitly encompasses conservatively modifiedvariants thereof (e.g., degenerate codon substitutions), alleles,orthologs, SNPs, and complementary sequences as well as the sequenceexplicitly indicated. Specifically, degenerate codon substitutions maybe achieved by generating sequences in which the third position of oneor more selected (or all) codons is substituted with mixed-base and/ordeoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991);Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); and Rossolini etal., Mol. Cell. Probes 8:91-98 (1994)) The nucleotide in the“polynucleotide” or “nucleic acid” may comprise modifications includingbase modifications such as bromouridine and inosine derivatives, ribosemodification such as phosphorothioate, phosphorodithioate,phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate,phosphoraniladate and phosphoroamidate.

The term “vector” means any molecule or entity (e.g. nucleic acid,plasmid, bacteriophage or virus) that is suitable for transformation ortransfection of a host cell and contains nucleic acid sequences thatdirect and/or control (in conjunction with the host cell) expression ofone or more heterologous coding regions operatively linked thereto.

A “conservative variant” of a sequence encoding a binding molecule, anantibody or a fragment thereof refers to a sequence comprisingconservative amino acid modifications. “Conservative amino acidmodifications” are intended to refer to amino acid modifications that donot significantly affect or alter the binding characteristics of theantibody containing the amino acid sequence. Such conservativemodifications include amino acid substitutions, additions and deletions.Conservative amino acid substitutions are ones in which the amino acidresidue is replaced with an amino acid residue having a similar sidechain. Families of amino acid residues having similar side chains havebeen defined in the art. These families include amino acids with basicside chains (e.g., lysine, arginine, histidine), acidic side chains(e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g.,glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine,tryptophan), nonpolar side chains (e.g., alanine, valine, leucine,isoleucine, proline, phenylalanine, methionine), beta-branched sidechains (e.g., threonine, valine, isoleucine) and aromatic side chains(e.g., tyrosine, phenylalanine, tryptophan, histidine). Modificationscan be introduced into a binding protein of the invention by standardtechniques known in the art, such as site-directed mutagenesis andPCR-mediated mutagenesis. Conservative amino acid substitution can alsoencompass non-naturally occurring amino acid residues which aretypically incorporated by chemical peptide synthesis rather than bysynthesis in biological systems. Non-naturally occurring amino acidsinclude, but are not limited to, peptidomimetic, reversed or invertedforms of amino acid moieties.

The term “epitope” is the part of an antigen that is recognized by theimmune system, such as an antibody or a fragment thereof. Within thepresent specification, the term “epitope” is used interchangeably forboth conformational epitopes and linear epitopes. A conformationalepitope is composed of discontinuous sections of the antigen's aminoacid sequence, whilst a linear epitope is formed by a continuoussequence of amino acids from the antigen.

The term “treat”, “treating”, “treatment”, “prevent”, “preventing” or“prevention” includes therapeutic treatments, prophylactic treatmentsand applications in which one reduces the risk that a subject willdevelop a disorder or other risk factor. Treatment does not require thecomplete curing of a disorder and encompasses the reduction of thesymptoms or underlying risk factors.

2. Binding Molecules

The term “binding molecule” as used herein means any protein or peptidethat binds specifically to the IL-18 polypeptide. “Binding molecule”includes, but it is not limited to, antibodies and fragments thereof,such as immunologically functional fragments. The term “immunologicallyfunctional fragment” of an antibody or immunoglobulin chain as usedherein is a species of binding protein comprising a portion (regardlessof how that portion is obtained or synthesized) of an antibody thatlacks at least some of the amino acids present in a full-length chainbut which is still capable of specifically binding the IL-18polypeptide. Such fragments are biologically active in that they bindthe IL-18 polypeptide. “Binding molecule” refers to proteins whichspecifically bind the IL-18 polypeptide and which might additionallyneutralize the interaction of the IL-18 polypeptide with the IL-18receptor.

The term “binding molecule” as used herein also excludes the naturallyoccurring IL-18 binding protein and the isolated IL-18BP, for instanceas described in WO2001/085201.

The binding molecule of the present invention specifically binds IL-18,wherein the binding molecule does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and wherein the binding molecule is notIL-18BP.

As used herein, the term “does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex” is intended to refer to a binding molecule thatbinds to the IL-18/IL-18 binding protein (IL-18 BP) complex with a K_(D)of 1×10⁻⁵M or greater.

One way to assess whether a binding molecule binds IL-18 but does notbind the IL-18/IL-18 binding protein (IL-18 BP) complex is describedherein in the Exemplification, section 9.

In another aspect of the present invention, the binding moleculespecifically binds IL-18, wherein the binding molecule does not competewith IL-18 binding protein (IL-18 BP) for binding to IL-18 when IL-18BPis bound to IL-18.

The terms “compete”, “competing” and “cross-compete” and grammaticalvariations thereof are used interchangeably herein to mean the abilityof a binding molecule to compete with the IL-18BP for binding IL-18 whenthe IL-18BP is already bound to IL-18. The binding molecules of theinvention, such as an antibody or a fragment thereof do not compete inthis sense. Competition between binding molecules is determined by anassay in which the binding molecule is tested for specific binding toIL-18 when IL-18 is bound to IL-18BP. For the avoidance of any doubt, ifa binding molecule can displace IL18 from an IL-18/IL-18BP complex, thenthat binding molecule competes with the IL-18 BP for binding IL-18.

The binding molecule according to the invention is not IL-18BP, eitherisolated or naturally occurring IL-18BP.

The binding molecule may be selected from the following scaffolds: anantibody, a fragment of an antibody, a single variable domain antibody,a bi- or multi-specific antibody, a multivalent antibody, a dualvariable domain antibody, an immuno-conjugate, a fibronectin molecule,an adnectin, a DARPin, an avimer, an affibody, an anticalin, an affilin,a protein epitope mimetic or combinations thereof and as describedherein below.

Preferably the IL-18 comprises from amino acid 37 to amino acid 193 ofSEQ ID NO:1 (human IL-18) or SEQ ID NO:2 (cynomolgus monkey IL-18).

IL-18BP structure is characterized by a single Ig-like domain andresembles the extracellular segment of cytokine receptors with Ig-likestructures. Human IL-18BP has been identified in four differentisoforms. IL-18BP isoform a (IL-18BPa) exhibited the greatest affinityfor IL-18 with a rapid on-rate, a slow off-rate, and a dissociationconstant (K_(D)) of 399 pM. IL-18BP isoform c (IL-18BPc) shares the Igdomain of IL-18BPa except for the last 29 amino acids at the C-terminus.The K_(D) of IL-18BPc is 10-fold less (2.94 nM) than IL-18BPa.Nevertheless, IL-18BPa and IL-18BPc neutralize IL-18 >95% at a molarexcess of two. IL-18BP isoforms b and d lack a complete Ig domain andlack the ability to bind or neutralize IL-18. Murine IL-18BP is known intwo isoforms, c and d isoforms, possessing identical Ig domains and alsoneutralizing >95% murine IL-18 at a molar excess of two. However, murineIL-18BPd, which shares a common C-terminal motif with human IL-18BPa,also neutralizes human IL-18 (Kim et al. (2000) Proc Natl Acad Sci,97(3):1190-1195).

Throughout this specification, the term “IL-18BP” refers to human,murine or viral IL-18 binding proteins in every isoform, whethernaturally occurring, isolated or engineered such as the IL-18BPdisclosed in WO2001/085201 which describes analogues of IL-18BP(“muteins”) wherein one or more amino acids are inserted, replaced bydifferent conservative substitutions or deleted, IL-18BP fused protein(e.g. fused protein of an IL-18BP and an immunoglobulin heavy chainregion or Fe) and functional derivatives such as PEG-ylated IL-18BP.

In one embodiment, the binding molecule according to the invention,which binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the binding molecule is not IL-18BP,wherein the binding molecule binds with an IL-18 epitope on IL-18 asdefined with reference to SEQ ID NO:1, wherein the epitope:

-   -   a. is comprised within the following amino acids of IL-18 as        defined with reference to SEQ ID NO:1:        -   i. amino acids 41 and 42 and amino acids 87 to 97; or        -   ii. amino acids 138 to 160; or        -   iii. amino acids 177 to 181; or        -   iv. amino acids 41 and 42, amino acids 87 to 97, amino acids            138 to 160 and amino acids 177 to 181; or        -   v. amino acids 41, 42, 87; 89; 90; or        -   vi. amino acids 93, 94; 95, 96; or        -   vii. amino acids 140; 141; 150; 177; or        -   viii. amino acids 92; 93; 94; 138; 140; 152; 157; or        -   ix. amino acids 142; 143; 150; 152; or        -   x. amino acids 143; 144; 145; 177; 180; or        -   xi. amino acids 41, 42, 87; 89; 90; 93, 94; 95, 96; 140;            141; 150; 177; or        -   xii. amino acids 92; 93; 94; 138; 140; 142; 143; 144; 145;            150; 152; 157; 177; 180; or        -   xiii. amino acids 41; 42, 87; 89; 90; 92; 93, 94; 95, 96;            138; 140; 141; 142; 143; 144; 145; 150; 152; 157; 177; 180;            or    -   b. comprises of at least one, two, three, four of the amino        acids as defined in any one of the groups (i) to (xiii) listed        in a); or    -   c. comprises the amino acids as defined in any one of the        groups (iv) to (xii) listed in a).

In another embodiment, the binding molecule according to the invention,which binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the binding molecule is not IL-18BP,wherein the binding molecule binds with an IL-18 epitope on IL-18 asdefined with reference to SEQ ID NO:1, wherein the epitope comprisesamino acids Arg140 and Glu152. In one embodiment the epitope furthercomprises any one or more of amino acids Gln92, Pro93, Gly95, Pro143,Glu157 or Glu177. In another embodiment the epitope further comprisesany one or more of amino acids Lys89, Arg94, Met96, Phe138, Ser141,Gly144, His145, Asp146, Gln150 or Leu180.

In one embodiment of the present invention, the binding moleculespecifically binds IL-18, wherein the binding molecule does not bind theIL-18/IL-18 binding protein isoform a or isoform c (IL-18 BPa orIL-18BPc) complex, wherein the binding molecule is selected from: anantibody, a fragment of an antibody, a single variable domain antibody,a bi- or multi-specific antibody, a multivalent antibody, a dualvariable domain antibody, an immuno-conjugate, a fibronectin molecule,an adnectin, a DARPin, an avimer, an affibody, an anticalin, an affilin,a protein epitope mimetic or combinations thereof. Preferably the IL-18comprises from amino acid 37 to amino acid 193 of SEQ ID NO:1 (humanIL-18) or SEQ ID NO:2 (cynomolgus monkey IL-18). More preferably thebinding molecule is an antibody or a fragment thereof.

In another embodiment, the binding molecule according to the invention,which binds IL-18 and does not bind the IL-18/IL-18 binding proteinisoform a or isoform c (IL-18 BPa or IL-18BPc) complex and wherein thebinding molecule is not IL-18BP, wherein the binding molecule binds withan IL-18 epitope on IL-18 as defined with reference to SEQ ID NO:1,wherein the epitope comprises amino acids Arg140 and Glu152. In oneembodiment the epitope further comprises any one or more of amino acidsGln92, Pro93, Gly95, Pro143, Glu157 or Glu177 and wherein the bindingmolecule is selected from: an antibody, a fragment of an antibody, asingle variable domain antibody, a bi- or multi-specific antibody, amultivalent antibody, a dual variable domain antibody, animmuno-conjugate, a fibronectin molecule, an adnectin, a DARPin, anavimer, an affibody, an anticalin, an affilin, a protein epitope mimeticor combinations thereof.

In another embodiment, the binding molecule according to the invention,which binds IL-18 and does not bind the IL-18/IL-18 binding proteinisoform a or isoform c (IL-18 BPa or IL-18BPc) complex and wherein thebinding molecule is not IL-18BP, wherein the binding molecule binds withan IL-18 epitope on IL-18 as defined with reference to SEQ ID NO:1,wherein the epitope comprises amino acids Arg140 and Glu152 and whereinthe binding molecule is an antibody or a fragment thereof. In thisembodiment the epitope may further comprises any one or more of aminoacids Gln92, Pro93, Gly95, Pro143, Glu157 or Glu177.

The binding molecule of the invention is capable of inhibiting one ormore of IL-18 biological activities such as Th1 modulation; Th2modulation, NK-modulation, neutrophil modulation, monocyte-macrophagelineage modulation, eosinophil modulation, B-cell modulation, cytokinemodulation, chemokine modulation; adhesion molecule modulation, and cellrecruitment modulation. In one embodiment the binding molecule of theinvention inhibits IL-18-dependent interferon gamma (INF-γ) production,preferably in KG-1 cells. In another embodiment of the invention, thebinding molecule inhibits IL-18-dependent interferon gamma (INF-γ)production in KG-1 cells with an IC₅₀ of 10 nM or less or of 1 nM orless or of 100 pM or less in an assay as defined herein below in theexamples.

The binding molecule of the invention that specifically binds IL-18 hasa dissociation constant (K_(D)) of 10 nM or less in an assay as definedherein below in the examples. In one embodiment the K_(D) is of 1 nM orless. In another embodiment the binding molecule has a K_(D) of 100 pMor less and the binding molecule is an antibody or a fragment thereof.

The binding molecule according to the invention can be a crystallizedbinding molecule, preferably a controlled release crystallized bindingmolecule and/or carrier-free. In one embodiment, the crystallizedbinding molecule is an antibody or a fragment thereof. In anotherembodiment the crystallized binding molecule has a greater half-life invivo than the soluble counterpart and retains its biological functionafter crystallization.

The binding molecule according to the invention can also be a part of acomplex which comprises the binding molecule and IL-18. Preferably thebinding molecule is an antibody or a fragment thereof in complex withIL-18.

Finally, the binding molecule according to the invention may alsocompete with murine antibody 125-2H for binding human IL-18.

2) Antibodies

Binding molecules according to the present invention include antibodiesor fragments thereof, isolated and structurally characterized asdescribed hereinafter and as shown in FIGS. 1 and 2.

2.1) Human Antibodies

As used herein, a human antibody or a fragment thereof comprises heavyor light chain variable regions or full length heavy or light chainsthat are “the product of” or “derived from” a particular germlinesequence if the variable regions or full length chains of the antibodyare obtained from a system that uses human germline immunoglobulingenes. Such systems include immunizing a transgenic mouse carrying humanimmunoglobulin genes with the antigen of interest or screening a humanimmunoglobulin gene library displayed on phage with the antigen ofinterest. A human antibody or fragment thereof that is “the product of”or “derived from” a human germline immunoglobulin sequence can beidentified as such by comparing the amino acid sequence of the humanantibody to the amino acid sequences of human germline immunoglobulinsand selecting the human germline immunoglobulin sequence that is closestin sequence (i.e., greatest % identity) to the sequence of the humanantibody. A human antibody that is “the product of” or “derived from” aparticular human germline immunoglobulin sequence may contain amino aciddifferences as compared to the germline sequence, due to, for example,naturally occurring somatic mutations or intentional introduction ofsite-directed mutation. However, a selected human antibody typically isat least 90% identical in amino acids sequence to an amino acid sequenceencoded by a human germline immunoglobulin gene and contains amino acidresidues that identify the human antibody as being human when comparedto the germline immunoglobulin amino acid sequences of other species(e.g., murine germline sequences). In certain cases, a human antibodymay be at least 60%, 70%, 80%, 90%, or at least 95%, or even at least96%, 97%, 98%, or 99% identical in amino acid sequence to the amino acidsequence encoded by the germline immunoglobulin gene. Typically, a humanantibody derived from a particular human germline sequence will displayno more than 10 amino acid differences from the amino acid sequenceencoded by the human germline immunoglobulin gene. In certain cases, thehuman antibody may display no more than 5, or even no more than 4, 3, 2,or 1 amino acid difference from the amino acid sequence encoded by thegermline immunoglobulin gene.

Human antibodies may be produced by a number of methods known to thoseof skill in the art. Human antibodies can be made by the hybridomamethod using human myeloma or mouse-human heteromyeloma cells lines(Kozbor, J Immunol; (1984) 133:3001; Brodeur, Monoclonal IsolatedAntibody Production Techniques and Applications, pp 51-63, Marcel DekkerInc, 1987). Alternative methods include the use of phage libraries ortransgenic mice both of which utilize human variable region repertories(Winter G; (1994) Annu Rev Immunol 12:433-455, Green L L, (1999) JImmunol Methods 231:11-23).

Several strains of transgenic mice are now available wherein their mouseimmunoglobulin loci has been replaced with human immunoglobulin genesegments (Tomizuka K, (2000) Proc Natl Acad Sci, 97:722-727; Fishwild DM (1996) Nature Biotechnol 14:845-851; Mendez M J, (1997) NatureGenetics 15:146-156). Upon antigen challenge such mice are capable ofproducing a repertoire of human antibodies from which antibodies ofinterest can be selected. Of particular note is the Trimera™ system(Eren R et al, (1988) Immunology 93:154-161) where human lymphocytes aretransplanted into irradiated mice, the Selected Lymphocyte Isolatedantibody System (SLAM, Babcook et al, Proc Natl Acad Sci (1996)93:7843-7848) where human (or other species) lymphocytes are effectivelyput through a massive pooled in vitro isolated antibody generationprocedure followed by deconvoluted, limiting dilution and selectionprocedure and the Xenomouse™ (Abgenix Inc). An alternative approach isavailable from Morphotek Inc using the Morphodoma™ technology.

Phage display technology can be used to produce human antibodies andfragments thereof, (McCafferty; (1990) Nature, 348:552-553 and GriffithsA D et al (1994) EMBO 13:3245-3260). According to this technique,isolated antibody variable domain genes are cloned in frame into eithera major or minor coat of protein gene of a filamentous bacteriophagesuch as M13 or fd and displayed (usually with the aid of a helper phage)as function isolated antibody fragments on the surface of the phageparticle. Selections based on the function properties of the isolatedantibody result in selection of the gene encoding the isolated antibodyexhibiting these properties. The phage display technique can be used toselect antigen specific antibodies from libraries made from human Bcells taken from individuals afflicted with a disease or disorder oralternatively from unimmunized human donors (Marks; J Mol Bio (1991)222:581-591). Where an intact human isolated antibody is desiredcomprising an Fc domain it is necessary redone the phage displayedderived fragment into a mammalian expression vectors comprising thedesired constant regions and establishing stable expressing cell lines.

The technique of affinity maturation (Marks; Biotechnol (1992)10:779-783) may be used to provide binding affinity wherein the affinityof the primary human isolated antibody is improved by sequentiallyreplacing the H and L chain variable regions with naturally occurringvariants and selecting on the basis of improved binding affinities.Variants of this technique such as ‘epitope imprinting’ are now alsoavailable (WO 93/06213; Waterhouse; Nucl Acids Res (1993) 21:2265-2266).

Various (enumerated) embodiments of the invention are described herein.It will be recognised that features specified in each embodiment may becombined with other specified features to provide further embodiments ofthe present invention.

Embodiment 1: The binding molecule which binds IL-18 and does not bindthe IL-18/IL-18 binding protein (IL-18 BP) complex wherein the bindingmolecule is an isolated human antibody or a fragment thereof;preferably, an isolated human monoclonal antibody or a fragment thereof.

Embodiment 2: The isolated human antibody or a fragment thereofaccording to embodiment 1 wherein the isolated human antibody or afragment thereof binds with an IL-18 epitope on IL-18 as defined withreference to SEQ ID NO:1, wherein the epitope:

-   -   a. is comprised within the following amino acids of IL-18 as        defined with reference to SEQ ID NO:1:        -   i. amino acids 41 and 42 and amino acids 87 to 97; or        -   ii. amino acids 138 to 160; or        -   iii. amino acids 177 to 181; or        -   iv. amino acids 41 and 42, amino acids 87 to 97, amino acids            138 to 160 and amino acids 177 to 181; or        -   v. amino acids 41, 42, 87; 89; 90; or        -   vi. amino acids 93, 94; 95, 96; or        -   vii. amino acids 140; 141; 150; 177; or        -   viii. amino acids 92; 93; 94; 138; 140; 152; 157; or        -   ix. amino acids 142; 143; 150; 152; or        -   x. amino acids 143; 144; 145; 177; 180; or        -   xi. amino acids 41, 42, 87; 89; 90; 93, 94; 95, 96; 140;            141; 150; 177; or        -   xii. amino acids 92; 93; 94; 138; 140; 142; 143; 144; 145;            150; 152; 157; 177; 180; or        -   xiii. amino acids 41; 42, 87; 89; 90; 92; 93, 94; 95, 96;            138; 140; 141; 142; 143; 144; 145; 150; 152; 157; 177; 180;            or    -   b. comprises of at least one, two, three, four of the amino        acids as defined in any one of the groups (i) to (xiii) listed        in a); or    -   c. comprises the amino acids as defined in any one of the        groups (iv) to (xii) listed in a).

Embodiment 3: The isolated human antibody or fragment thereof accordingto embodiment 1, which binds to an IL-18 epitope on IL-18 as definedwith reference to SEQ ID NO:1, wherein the epitope comprises amino acidsArg140 and Glu152.

Embodiment 4: The isolated human antibody or fragment thereof accordingto embodiment 3, wherein the epitope further comprises any one or moreof amino acids Gln92, Pro93, Gly95, Pro143, Glu157 or Glu177.

Embodiment 5: The isolated human antibody or fragment thereof accordingto embodiments 3 and 4, wherein the epitope further comprises any one ormore of amino acids Lys89, Arg94, Met96, Phe138, Ser141, Gly144, His145,Asp146, Gln150 or Leu180.

Embodiment 6: The isolated human antibody or fragment thereof accordingto anyone of the preceding embodiments, wherein IL-18 comprises fromamino acid 37 to amino acid 193 of SEQ ID NO:1 or SEQ ID NO:2.

Embodiment 7: The isolated human antibody or fragment thereof accordingto anyone of the preceding embodiments, wherein the isolated humanantibody or fragment thereof inhibits IL-18-dependent interferon gamma(INF-γ) production.

Embodiment 8: The isolated human antibody or fragment thereof accordingto anyone of the preceding embodiments, wherein the isolated humanantibody or fragment thereof binds IL-18 with a K_(D) of 100 pM or less.

Embodiment 9: The isolated human antibody or fragment thereof accordingto anyone of the preceding embodiments, wherein the isolated humanantibody or fragment thereof further compete with murine antibody 125-2Hfor binding IL-18.

Embodiment 10: The isolated human antibody or fragment thereof accordingto anyone of the preceding embodiments, wherein the isolated humanantibody or fragment thereof binds IL-18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex and comprises a heavychain variable region H-CDR1 comprising SEQ ID NO: 3 or conservativevariants thereof and a heavy chain variable region H-CDR2 comprising SEQID NO: 4 or SEQ ID NO: 9 or SEQ ID NO: 10 or SEQ ID NO: 11 or SEQ ID NO:12 or SEQ ID NO: 13 or conservative variants thereof and a heavy chainvariable region H-CDR3 comprising SEQ ID NO: 5 or conservative variantsthereof and a light chain variable region L-CDR1 comprising SEQ ID NO: 6or conservative variants thereof and a light chain variable regionL-CDR2 comprising SEQ ID NO: 7 or conservative variants thereof and alight chain variable region L-CDR3 comprising SEQ ID NO: 8 orconservative variants thereof.

Embodiment 11: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 4 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 12: The isolated human antibody or fragment thereof accordingto embodiment 11, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 4 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof and wherein theisolated human antibody or fragment thereof competes with murineantibody 125-2H for binding IL-18.

Embodiment 13: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 9 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 14: The isolated human antibody or fragment thereof accordingto embodiment 13, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 9 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof and wherein theisolated human antibody or fragment thereof competes with murineantibody 125-2H for binding IL-18.

Embodiment 15: The isolated human antibody or fragment thereof accordingto embodiments 13 or 14, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and that antibody or fragment thereof binds to anepitope comprising Arg140 and Glu152 on IL-18 as defined with referenceto SEQ ID NO:1, wherein the isolated antibody or fragment thereofcomprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO: 3        or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO: 9        or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        5 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO: 6        or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO: 7        or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO: 8        or conservative variants thereof.

Preferably this isolated human antibody is an isolated fully humanantibody or fragment thereof, more preferably an isolated fully humanmonoclonal antibody or fragment thereof.

Embodiment 16: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 10 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 17: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 11 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 18: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 12 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 19: The isolated human antibody or fragment thereof accordingto embodiment 10, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 13 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof.

Embodiment 20: The isolated human antibody or fragment thereof accordingto embodiment 19, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 13 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 5 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 7 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 8 or conservative variants thereof and wherein theisolated human antibody or fragment thereof competes with murineantibody 125-2H for binding IL-18.

Embodiment 21: The isolated human antibody or fragment thereof accordingto embodiments 19 or 20, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and that antibody or fragment thereof binds to anepitope comprising Arg140 and Glu152 on IL-18 as defined with referenceto SEQ ID NO:1 wherein the isolated antibody or fragment thereofcomprises:

-   -   i. a heavy chain variable region H-CDR1 comprising SEQ ID NO: 3        or conservative variants thereof and    -   ii. a heavy chain variable region H-CDR2 comprising SEQ ID NO:        13 or conservative variants thereof and    -   iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO:        5 or conservative variants thereof and    -   iv. a light chain variable region L-CDR1 comprising SEQ ID NO: 6        or conservative variants thereof and    -   v. a light chain variable region L-CDR2 comprising SEQ ID NO: 7        or conservative variants thereof and    -   vi. a light chain variable region L-CDR3 comprising SEQ ID NO: 8        or conservative variants thereof.

Preferably this isolated human antibody is an isolated fully humanantibody or fragment thereof, more preferably an isolated fully humanmonoclonal antibody or fragment thereof.

Embodiment 22: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprises a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 74 or conservative variants thereofand a heavy chain variable region H-CDR2 comprising SEQ ID NO: 75 or SEQID NO: 76 or SEQ ID NO: 77 or SEQ ID NO: 78 or conservative variantsthereof and a heavy chain variable region H-CDR3 comprising SEQ ID NO:79 or conservative variants thereof and a light chain variable regionL-CDR1 comprising SEQ ID NO: 80 or conservative variants thereof and alight chain variable region L-CDR2 comprising SEQ ID NO: 81 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 82 or conservative variants thereof.

Embodiment 23: The isolated human antibody or fragment thereof accordingto embodiment 22, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 74 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 75 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 79 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 80 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 81 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 82 or conservative variants thereof.

Embodiment 24: The isolated human antibody or fragment thereof accordingto embodiment 22, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 74 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 76 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 79 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 80 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 81 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 82 or conservative variants thereof.

Embodiment 25: The isolated human antibody or fragment thereof accordingto embodiment 22, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 74 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 77 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 79 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 80 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 81 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 82 or conservative variants thereof.

Embodiment 26: The isolated human antibody or fragment thereof accordingto embodiment 22, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 74 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 78 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 79 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 80 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 81 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 82 or conservative variants thereof.

Embodiment 27: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprises a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 106 or conservative variants thereofand a heavy chain variable region H-CDR2 comprising SEQ ID NO: 107 orSEQ ID NO: 122 or conservative variants thereof and a heavy chainvariable region H-CDR3 comprising SEQ ID NO: 108 or conservativevariants thereof and a light chain variable region L-CDR1 comprising SEQID NO: 109 or conservative variants thereof and a light chain variableregion L-CDR2 comprising SEQ ID NO: 110 or conservative variants thereofand a light chain variable region L-CDR3 comprising SEQ ID NO: 111 orSEQ ID NO: 126 or conservative variants thereof.

Embodiment 28: The isolated human antibody or fragment thereof accordingto embodiment 27, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 106 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 107 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 108 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 109 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 110 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 111 or conservative variants thereof.

Preferably this isolated human antibody or fragment thereof is anisolated fully human antibody or fragment thereof, more preferably anisolated fully human monoclonal antibody or fragment thereof.

Embodiment 29: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprises a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 106 or conservative variants thereofand a heavy chain variable region H-CDR2 comprising SEQ ID NO: 122 orconservative variants thereof and a heavy chain variable region H-CDR3comprising SEQ ID NO: 108 or conservative variants thereof and a lightchain variable region L-CDR1 comprising SEQ ID NO: 109 or conservativevariants thereof and a light chain variable region L-CDR2 comprising SEQID NO: 110 or conservative variants thereof and a light chain variableregion L-CDR3 comprising SEQ ID NO: 126 or conservative variantsthereof.

Embodiment 30: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprise a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 120 or conservative variants thereofand a heavy chain variable region H-CDR2 comprising SEQ ID NO: 121 orconservative variants thereof and a heavy chain variable region H-CDR3comprising SEQ ID NO: 123 or conservative variants thereof and a lightchain variable region L-CDR1 comprising SEQ ID NO: 124 or conservativevariants thereof and a light chain variable region L-CDR2 comprising SEQID NO: 125 or conservative variants thereof and a light chain variableregion L-CDR3 comprising SEQ ID NO: 127 or SEQ ID NO: 128 or SEQ ID NO:129 or conservative variants thereof.

Embodiment 31: The isolated human antibody or fragment thereof accordingto embodiment 30, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 120 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 121 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 123 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 124 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 125 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 127 or conservative variants thereof.

Embodiment 32: The isolated human antibody or fragment thereof accordingto embodiment 30, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 120 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 121 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 123 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 124 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 125 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 128 or conservative variants thereof.

Embodiment 33: The isolated human antibody or fragment thereof accordingto embodiment 30, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 120 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 121 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 123 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 124 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 125 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 129 or conservative variants thereof.

The CDR regions outlined above are delineated using the Kabat system(Kabat, E. A., et al., 1991 Sequences of Proteins of ImmunologicalInterest, Fifth Edition, U.S. Department of Health and Human Services,NIH Publication No. 91-3242).

In some other embodiments of the invention, the human antibody orfragment thereof comprises the CDR regions delineated using the Chothiadefinition (Chothia et al., (1987) J Mol Biol 196: 901-17).

Embodiment 34: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprises a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 59 or SEQ ID NO: 65 or SEQ ID NO: 66or conservative variants thereof and a heavy chain variable regionH-CDR2 comprising SEQ ID NO: 60 or SEQ ID NO: 67 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 61 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 62 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 63 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 64 or conservative variants thereof.

Embodiment 35: The isolated human antibody or fragment thereof accordingto embodiment 34, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 59 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 60 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 61 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 62 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 63 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 64 or conservative variants thereof.

Embodiment 36: The isolated human antibody or fragment thereof accordingto embodiment 35, wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 37: The isolated human antibody or fragment thereof accordingto embodiment 34, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 65 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 60 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 61 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 62 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 63 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 64 or conservative variants thereof.

Embodiment 38: The isolated human antibody or fragment thereof accordingto embodiment 37, wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 39: The isolated human antibody or fragment thereof accordingto embodiment 34, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region H-CDR1comprising SEQ ID NO: 66 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 67 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 61 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 62 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 63 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 64 or conservative variants thereof.

Embodiment 40: The isolated human antibody or fragment thereof accordingto embodiment 39, wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 41: The isolated human antibody or fragment thereof accordingto embodiment 34, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprise a heavy chain variable region H-CDR1comprising SEQ ID NO: 68 or conservative variants thereof and a heavychain variable region H-CDR2 comprising SEQ ID NO: 69 or conservativevariants thereof and a heavy chain variable region H-CDR3 comprising SEQID NO: 70 or conservative variants thereof and a light chain variableregion L-CDR1 comprising SEQ ID NO: 71 or conservative variants thereofand a light chain variable region L-CDR2 comprising SEQ ID NO: 72 orconservative variants thereof and a light chain variable region L-CDR3comprising SEQ ID NO: 73 or conservative variants thereof.

Embodiment 42: The isolated human antibody or fragment thereof accordingto anyone of the embodiments 1 to 9, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and comprise a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 162 or conservative variants thereofand a heavy chain variable region H-CDR2 comprising SEQ ID NO: 163 orconservative variants thereof and a heavy chain variable region H-CDR3comprising SEQ ID NO: 164 or conservative variants thereof and a lightchain variable region L-CDR1 comprising SEQ ID NO: 165 or conservativevariants thereof and a light chain variable region L-CDR2 comprising SEQID NO: 166 or conservative variants thereof and a light chain variableregion L-CDR3 comprising SEQ ID NO: 167 or conservative variantsthereof.

Under Kabat definition, as discussed above, the CDR amino acid residuesin the heavy chain variable domain (VH) are numbered 31-35 (H-CDR1),50-65 (H-CDR2), and 95-102 (H-CDR3); and the CDR amino acid residues inthe light chain variable domain (VL) are numbered 24-34 (L-CDR1), 50-56(L-CDR2), and 89-97 (L-CDR3). Under Chothia the CDR amino acids in theVH are numbered 26-32 (H-CDR1), 52-56 (H-CDR2), and 95-102 (H-CDR3); andthe amino acid residues in VL are numbered 26-32 (L-CDR1), 50-52(L-CDR2), and 91-96 (L-CDR3). By combining the CDR definitions of bothKabat and Chothia and taking into consideration insertion for longerloops, the CDRs consist of amino acid residues 26-35 (H-CDR1), 50-65(H-CDR2), and 95-102 (H-CDR3) in human VH and amino acid residues 24-34(L-CDR1), 50-56 (L-CDR2), and 89-97 (L-CDR3) in human VL.

Embodiment 43: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) comprising SEQ ID NO: 14 or conservative variants thereof and alight chain variable region (VL) comprising SEQ ID NO: 16 orconservative variants thereof and wherein the heavy chain variableregion (VH) comprises:

-   -   i. a heavy chain variable region H-CDR1 corresponding to amino        acids 26 to 35 SEQ ID NO: 14; and    -   ii. a heavy chain variable region H-CDR2 corresponding to amino        acids 50 to 66 SEQ ID NO: 14; and    -   iii. a heavy chain variable region H-CDR3 corresponding to amino        acids 99 to 108 SEQ ID NO: 14;

and wherein the light chain variable region (VL) comprises:

-   -   iv. a light chain variable region L-CDR1 corresponding to amino        acids 23 to 35 SEQ ID NO: 16; and    -   v. a light chain variable region L-CDR2 corresponding to amino        acids 51 to 57 SEQ ID NO: 16; and    -   vi. a light chain variable region L-CDR3 corresponding to amino        acids 90 to 100 SEQ ID NO: 16.

Embodiment 44: The isolated human antibody or fragment thereof accordingto embodiment 43, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 45: The isolated human antibody or fragment thereof accordingto embodiments 43 or 44, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and that antibody or fragment thereof binds to anepitope comprising Arg140 and Glu152 on IL-18 as defined with referenceto SEQ ID NO:1, optionally the epitope further comprises any one or moreof amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96, Phe138, Ser141,Pro143, Gly144, Glu157, Glu177 or Leu180, preferably the epitope furthercomprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96, Phe138,Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 46: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) comprising SEQ ID NO: 18 or conservative variants thereof and alight chain variable region (VL) comprising SEQ ID NO: 20 orconservative variants thereof and wherein the heavy chain variableregion (VH) comprises:

-   -   i. a heavy chain variable region H-CDR1 corresponding to amino        acids 26 to 35 SEQ ID NO: 18; and    -   ii. a heavy chain variable region H-CDR2 corresponding to amino        acids 50 to 66 SEQ ID NO: 18; and    -   iii. a heavy chain variable region H-CDR3 corresponding to amino        acids 99 to 108 SEQ ID NO: 18;

and wherein the light chain variable region (VL) comprises:

-   -   iv. a light chain variable region L-CDR1 corresponding to amino        acids 23 to 35 SEQ ID NO: 20; and    -   v. a light chain variable region L-CDR2 corresponding to amino        acids 51 to 57 SEQ ID NO: 20; and    -   vi. a light chain variable region L-CDR3 corresponding to amino        acids 90 to 100 SEQ ID NO: 20.

Embodiment 47: The isolated human antibody or fragment thereof accordingto embodiment 46, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 48: The isolated human antibody or fragment thereof accordingto embodiments 46 or 47, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and that antibody or fragment thereof binds to anepitope comprising Arg140 and Glu152 on IL-18 as defined with referenceto SEQ ID NO:1.

Given that each of these human antibodies can bind to IL18 and thatantigen-binding specificity is provided primarily by the CDR1, 2 and 3regions, the H-CDR1, 2 and 3 sequences and L-CDR1, 2 and 3 sequences canbe “mixed and matched” (i.e., CDRs from different human antibodies canbe mixed and matched, each antibody containing a H-CDR1, 2 and 3 set anda L-CDR1, 2 and 3 set create other anti-IL18 binding molecules of theinvention. IL18 binding of such “mixed and matched” antibodies can betested using the binding assays in the Examples (e.g., ELISAs). When VHCDR sequences are mixed and matched, the CDR1, CDR2 and/or CDR3 sequencefrom a particular VH sequence should be replaced with a structurallysimilar CDR sequence(s). Likewise, when VL CDR sequences are mixed andmatched, the CDR1, CDR2 and/or CDR3 sequence from a particular VLsequence should be replaced with a structurally similar CDR sequence(s).It will be readily apparent to the ordinarily skilled artisan that novelVH and VL sequences can be created by substituting one or more VH and/orVL CDR region sequences with structurally similar sequences from the CDRsequences shown herein for human antibodies of the present invention(FIGS. 1 and 2).

In another aspect, the invention provides an isolated human antibodybinding IL-18 and not binding the IL-18/IL-18 binding protein (IL-18 BP)complex and comprising VH amino acid sequences selected from thesequences shown in SEQ ID NOs: 14, 18, 22, 25, 28, 31, 34, 37, 40, 83,87, 90, 93, 112, 130 and 138 and VL amino acid sequences selected fromthe sequences shown in SEQ ID NOs: 16, 20, 85, 114, 132, 140, 147 and153. Other antibodies of the invention include amino acids that havebeen mutated by amino acid deletion, insertion or substitution, yet haveat least 60, 70, 80, 90 or 95 percent identity in the CDR regions withthe CDR regions depicted in the sequences described above. In someembodiments, it include mutant amino acid sequences wherein no more than1, 2, 3, 4 or 5 amino acids have been mutated by amino acid deletion,insertion or substitution in the CDR regions when compared with the CDRregions depicted in the sequences described above.

Embodiment 49: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequence shown in SEQ ID NO:28 or conservative variants thereof and a light chain variable region(VL) amino acid sequence selected from the sequence shown in SEQ ID NO:16 or conservative variants thereof.

Embodiment 50: The isolated human antibody or fragment thereof accordingto embodiment 49, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 28 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 16 orconservative variants thereof, wherein amino acid asparagine (Asn; N) inposition 30 with reference to SEQ ID NO: 28 is replaced by an amino acidselected from lysine (Lys; K) or serine (Ser; S) or threonine (Thr; T)or alanine (Ala; A) or glutamate (Glu; E) or histidine (His; H) orleucine (Leu; L) or glutamine (Gln; Q) or arginine (Arg; R) or valine(Val; V) or tyrosine (Tyr; Y) or isoleucine (Ile; I) or glycine (Gly;G).

Embodiment 51: The isolated human antibody or fragment thereof accordingto embodiments 49 or 50, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 52: The isolated human antibody or fragment thereof accordingto any one of embodiments 49 to 51, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 53: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequence shown in SEQ ID NO:14 or conservative variants thereof and a light chain variable region(VL) amino acid sequence selected from the sequence shown in SEQ ID NO:16 or conservative variants thereof.

Embodiment 54: The isolated human antibody or fragment thereof accordingto embodiment 53, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 55: The isolated human antibody or fragment thereof accordingto any one of embodiments 53 to 55, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 56: The isolated human antibody or fragment thereof accordingto embodiment 53, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 14 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 16 orconservative variants thereof, wherein amino acid lysine (Lys; K) inposition 30 with reference to SEQ ID NO: 14 is replaced by an amino acidselected from asparagine (Asn; N) or serine (Ser; S) or threonine (Thr;T) or alanine (Ala; A) or glutamate (Glu; E) or histidine (His; H) orleucine (Leu; L) or glutamine (Gln; Q) or arginine (Arg; R) or valine(Val; V) or tyrosine (Tyr; Y) or isoleucine (Ile; I) or glycine (Gly;G).

Embodiment 57: The isolated human antibody or fragment thereof accordingto embodiment 56, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 58: The isolated human antibody or fragment thereof accordingto any one of embodiments 55 or 57, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 59: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequences shown in SEQ ID NO:18 or conservative variants thereof and a light chain variable region(VL) amino acid sequence selected from the sequence shown in SEQ ID NO:20 or conservative variants thereof.

Embodiment 60: The isolated human antibody or fragment thereof accordingto embodiment 59, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 61: The isolated human antibody or fragment thereof accordingto any one of embodiments 59 or 60, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 62: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequences shown in SEQ ID NO:40 or conservative variants thereof and a light chain variable region(VL) amino acid sequence selected from the sequence shown in SEQ ID NO:20 or conservative variants thereof.

Embodiment 63: The isolated human antibody or fragment thereof accordingto embodiment 62, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 40 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 20 orconservative variants thereof, wherein

-   -   i. amino acid glutamate (Glu; E) in position 1 with reference to        SEQ ID NO: 40 is replaced by amino acid glutamine (Gln; Q) and    -   ii. wherein amino acid asparagine (Asn; N) in position 30 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from serine (Ser; S) or threonine (Thr; T) or aspartate (Asp;        D).

Embodiment 64: The isolated human antibody or fragment thereof accordingto embodiment 63, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 40 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 20 orconservative variants thereof, wherein

-   -   i. amino acid glutamate (Glu; E) in position 1 with reference to        SEQ ID NO: 40 is replaced by amino acid glutamine (Gln; Q); and    -   ii. wherein amino acid asparagine (Asn; N) in position 30 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from serine (Ser; S) or threonine (Thr; T) or aspartate (Asp;        D); and    -   iii. wherein amino acid methionine (Met; M) in position 54 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from tyrosine (Tyr; Y) or asparagine (Asn; N) or isoleucine        (Ile; I).

Embodiment 65: The isolated human antibody or fragment thereof accordingto embodiments 62 to 64, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 66: The isolated human antibody or fragment thereof accordingto any one of embodiments 65 to 66, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 67: The isolated human antibody or fragment thereof accordingto embodiment 62, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 40 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 20 orconservative variants thereof, wherein

-   -   i. amino acid glutamate (Glu; E) in position 1 with reference to        SEQ ID NO: 40 is replaced by amino acid glutamine (Gln; Q) and    -   ii. wherein amino acid serine (Ser; S) in position 31 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from threonine (Thr; T) or asparagine (Asn; N) or alanine (Ala;        A).

Embodiment 68: The isolated human antibody or fragment thereof accordingto embodiment 67, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and comprises a heavy chain variable region (VH)amino acid sequence selected from the sequence shown in SEQ ID NO: 40 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 20 orconservative variants thereof, wherein

-   -   i. amino acid glutamate (Glu; E) in position 1 with reference to        SEQ ID NO: 40 is replaced by amino acid glutamine (Gln; Q); and    -   ii. wherein amino acid serine (Ser; S) in position 31 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from threonine (Thr; T) or asparagine (Asn; N) or alanine (Ala;        A).    -   iii. wherein amino acid methionine (Met; M) in position 54 with        reference to SEQ ID NO: 40 is replaced by an amino acid selected        from tyrosine (Tyr; Y) or asparagine (Asn; N) or isoleucine        (Ile; I).

Embodiment 69: The isolated human antibody or fragment thereof accordingto embodiments 67 or 68, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 70: The isolated human antibody or fragment thereof accordingto any one of embodiments 67 to 69, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 72: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequences shown in SEQ ID NO:22 or SEQ ID NO: 25 SEQ ID NO: 28 or SEQ ID NO: 31 or SEQ ID NO: 34 orconservative variants thereof and a light chain variable region (VL)amino acid sequence selected from the sequence shown in SEQ ID NO: 16 orconservative variants thereof.

Embodiment 73: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a heavy chain variable region(VH) amino acid sequence selected from the sequences shown in SEQ ID NO:37 and a light chain variable region (VL) amino acid sequence selectedfrom the sequence shown in SEQ ID NO: 20 or conservative variantsthereof.

Embodiment 74: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequences shown in SEQ ID NOs: 43, 47, 50, 53, 56, 96,100, 103, 116, 134, 142 and 158 or conservative variants thereof; and aVL amino acid sequence selected from the sequences shown in SEQ ID NOs:45, 98, 118, 136, 144, 150, 156 and 160 or conservative variantsthereof.

Embodiment 75: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequence shown in SEQ ID NO: 43 or conservativevariants thereof and a VL amino acid sequence selected from the sequenceshown in SEQ ID NO: 45 or conservative variants thereof.

Embodiment 76: The isolated human antibody or fragment thereof accordingto embodiment 75, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 77: The isolated human antibody or fragment thereof accordingto any one of embodiments 75 or 76, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 78: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequence shown in SEQ ID NO: 158 or conservativevariants thereof and a VL amino acid sequence selected from the sequenceshown in SEQ ID NO: 160 or conservative variants thereof.

Embodiment 79: The isolated human antibody or fragment thereof accordingto embodiment 78, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 80: The isolated human antibody or fragment thereof accordingto any one of embodiments 78 or 79, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 81: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequences shown in SEQ ID NO: 47 or SEQ ID NO: 50 orSEQ ID NO: 56 or conservative variants thereof and a VL amino acidsequence selected from the sequence shown in SEQ ID NO: 45 orconservative variants thereof.

Embodiment 82: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequences shown in SEQ ID NO: 53 or SEQ ID NO: 100 orconservative variants thereof and a VL amino acid sequence selected fromthe sequences shown in SEQ ID NO: 160 or conservative variants thereof.

Embodiment 83: The isolated human antibody or fragment thereof accordingto embodiments 81 or 82, wherein the isolated human antibody or fragmentthereof binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex and wherein the isolated human antibody or fragmentthereof competes with murine antibody 125-2H for binding IL-18.

Embodiment 84: The isolated human antibody or fragment thereof accordingto any one of embodiments 81 to 83, wherein the isolated human antibodyor fragment thereof binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and that antibody or fragment thereofbinds to an epitope comprising Arg140 and Glu152 on IL-18 as definedwith reference to SEQ ID NO:1, optionally the epitope further comprisesany one or more of amino acids Lys89, Gln92, Pro93, Arg94, Gly95, Met96,Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 or Leu180, preferably theepitope further comprises amino acids Lys89, Gln92, Pro93, Arg94, Gly95,Met96, Phe138, Ser141, Pro143, Gly144, Glu157, Glu177 and Leu180.

Embodiment 85: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequences shown in SEQ ID NO: 96 or SEQ ID NO: 103 orconservative variants thereof and a VL amino acid sequence selected fromthe sequences shown in SEQ ID NO: 98 or conservative variants thereof.

Embodiment 86: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequence shown in SEQ ID NO: 116 or conservativevariants thereof and a VL amino acid sequence selected from the sequenceshown in SEQ ID NO: 118 or conservative variants thereof.

Embodiment 87: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequence shown in SEQ ID NO: 142 or conservativevariants thereof and a VL amino acid sequence selected from the sequenceshown in SEQ ID NO: 144 or conservative variants thereof.

Embodiment 88: The isolated human antibody or fragment thereof accordingto any one of embodiments 1 to 9, wherein the isolated human antibody orfragment thereof binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex and comprises a VH amino acid sequenceselected from the sequence shown in SEQ ID NO: 134 or conservativevariants thereof and a VL amino acid sequence selected from thesequences shown in SEQ ID NO: 136 or SEQ ID NO: 150 or SEQ ID NO: 156 orconservative variants thereof.

2.2) Humanized or Chimeric Antibodies

An obvious alternative to the invention disclosed herein is the use ofhumanized or chimeric antibodies in place of human antibodies.

The use of intact non-human antibodies in the treatment of humandiseases or disorders carries with it the potential for the now wellestablished problems of immunogenicity. That is, the immune system ofthe patient may recognise the non-human intact isolated antibody asnon-self and mount an antibody response. This is particularly evidentupon multiple administration of the non-human isolated antibody to ahuman patient. Various techniques have been developed over the years toovercome these problems and generally involve reducing the non-humanimmunogenicity signature in the intact isolated antibody whilstretaining the relative ease in obtaining non-human antibodies from animmunised animal, e.g. mouse, rat or rabbit. Broadly two approaches havebeen used to achieve this. The first are chimeric (sometimes“chimaeric”) antibodies, which generally comprise a non-human (e.g.rodent such as mouse) variable domain fused to a human constant region.Because the antigen-binding site of an isolated antibody is localisedwithin the variable regions the chimeric isolated antibody retains itsbinding affinity for the antigen but acquires the effector functions ofthe human constant region and are therefore able to perform effectorfunctions such as described supra. Chimeric antibodies are typicallyproduced using recombinant DNA methods. DNA encoding the antibodies(e.g. cDNA) is isolated and sequenced using conventional procedures(e.g. by using oligonucleotide probes that are capable of bindingspecifically to genes encoding the heavy and light chains of theisolated antibody of the invention. Hybridoma cells serve as a typicalsource of such DNA. Once isolated, the DNA is placed into expressionvectors which are then transfected into host cells such as E. coli, COScells, CHO cells or myeloma cells that do not otherwise produceimmunoglobulin protein to obtain synthesis of the isolated antibody. TheDNA may be modified by substituting the coding sequence for human L andH chains for the corresponding non-human (e.g. murine) H and L constantregions (Morrison; PNAS 81, 6851 (1984)).

The second approach involves the generation of humanized antibodieswherein the non-human content of the isolated antibody is reduced byhumanizing the variable regions. Two techniques for humanization havegained popularity. The first is humanization by CDR grafting. CDRs buildloops close to the isolated antibody's N-terminus where they form asurface mounted in a scaffold provided by the framework region.Antigen-binding specificity of the isolated antibody is mainly definedby the topography and by the chemical characteristics of its CDRsurface. These features are in turn determined by the conformation ofthe individual CDRs, by the relative disposition of the CDRs, and by thenature and disposition of the side chains of the residues comprising theCDRs. A large decrease in immunogenicity can be achieved by graftingonly the CDRs of a non-human (e.g. murine) antibodies (‘donor’antibodies) onto human framework (‘acceptor framework’) and constantregions (Jones et al (1986) Nature 321:522-525 and Verhoeyen M et al(1988) Science 239:1534-1536). However, CDR grafting per se may notresult in the complete retention of antigen-binding properties and it isfrequency found that some framework residues (sometimes referred to as‘backmutations’) of the donor isolated antibody need to be preserved inthe humanised compound if significant antigen-binding affinity is to berecovered (Queen C et al., (1989) Proc Natl Acad Sci 86:10029-10033, Co,M et al (1991) Nature 351, 501-502). In this case, human variableregions showing the greatest sequence homology to the non-human donorisolated antibody are chosen from a database in order to provide thehuman framework (FR). The selection of human FRs can be made either fromhuman consensus or individual human antibodies. Where necessary, keyresidues from the donor isolated antibody are substituted into the humanacceptor framework to preserve CDR conformations. Computer modelling ofthe isolated antibody may be used to help identify such structurallyimportant residues.

Alternatively, humanisation may be achieved by a process of ‘veneering’.A statistical analysis of unique human and murine immunoglobulin heavyand light chain variable regions revealed that the precise patterns ofexposed residues are different in human and murine antibodies, and mostindividual surface positions have a strong preference for a small numberof different residues (Padlan E A, et al; (1991) Mol Immunol 28:489-498and Pedersen J T et al (1994) J Mol Biol 235:959-973). Therefore it ispossible to reduce the immunogenicity of a non-human Fv by replacingexposed residues in its framework regions that differ from those usuallyfound in human antibodies. Because protein antigenicity may becorrelated with surface accessibility, replacement of the surfaceresidues may be sufficient to render the mouse variable region‘invisible’ to the human immune system (also Mark G E et al (1994) inHandbook of Experimental Pharmacology vol 113: The pharmacology ofmonoclonal Antibodies, Springer-Verlag, pp 105 134). This procedure ofhumanisation is referred to as ‘veneering’ because only the surface ofthe isolated antibody is altered, the supporting residues remainundisturbed.

2.3) Isolated Antibody Fragments and Single Variable Domain Antibodies

In another embodiment of the invention, the binding molecule is afragment of an antibody or a single variable domain antibody which bindsIL-18 and does not bind the IL-18/IL-18 binding protein (IL-18 BP)complex. Preferably, the binding molecule is a Fab, a Fab′, a F(ab′)₂, aFv, a scFv, a dAb or a V_(HH).

Traditionally, such fragments are produced by the proteolytic digestionof intact antibodies by e.g. papain digestion (see for example WO94/29348) but may be produced directly from recombinantly transformedhost cells. In addition, isolated antibody fragments may be producedusing a variety of engineering techniques as described below.

Fv fragments appear to have lower interaction energy of their two chainsthan Fab fragments. To stabilise the association of the VH and VLdomains, they have been linked with peptides (Bird et al, (1988)Science, 242:423-426, Huston et al, (1998) PNAS, 85:5879-5883),disulphide bridges (Glockshuber et al, (1990) Biochemistry,29:1362-1367) and ‘knob in hole’ mutations (Zhu et al (1997), ProteinSci, 6:781-788). ScFv fragments can be produced by methods well known tothose skilled in the art (Whitlow et al (1991), Methods companionMethods Enzymol, 2:97-105 and Huston et al (1993) Int Rev Immunol10:195-217). ScFv may be produced in bacterial cells such as E. coli butare more preferably produced in eukaryotic cells. One disadvantage ofscFv is the monovalency of the product, which precludes an increasedavidity due to polyvalent binding, and their short half-life. Attemptsto overcome these problems include bivalent (scFv′)₂ produced from scFvcontaining an additional C terminal cysteine by chemical coupling (Adamset al (1993) Can Res 53:4026-4034 and McCartney et al (1995) ProteinEng, 8:301-314) or by spontaneous site-specific dimerization of scFvcontaining an unpaired C terminal cysteine residue (Kipriyanov et al(1995) Cell. Biophys 26:187-204). Alternatively, scFv can be forced toform multimers by shortening the peptide linker to 3 and 12 residues toform ‘diabodies’ (Holliger et al PNAS (1993), 90:6444-6448). Reducingthe linker still further can result in scFv trimers (‘triabodies’, seeKortt et al (1997) Protein Eng, 10:423-433) and tetramers(‘tetrabodies’, Le Gall et al (1999) FEBS Lett, 453:164-168).Construction of bivalent scFv compounds can also be achieved by geneticfusion with protein dimerzing motifs to form ‘miniantibodies’ (Pack etal (1992) Biochemistry 31:1579-1584) and ‘minibodies’ (Hu et al (1996),Cancer Res. 56:3055-3061). ScFv-sc-Fv tandems ((scFv)2) may also beproduced by linking two scFv units by a third peptide linker, (seeKurucz et al (1995) J Immunol, 154:4576-4582). Bispecific diabodies canbe produced through the noncovalent association of two single chainfusion products consisting of VH domain from one isolated antibodyconnected by a short linker to the VL domain of another isolatedantibody, (see Kipriyanov et al (1998), Int J Can 77:763-772). Thestability of such bispecific diabodies can be enhanced by theintroduction of disulphide bridges or ‘knob in hole’ mutations asdescribed supra or by the formation of single chain diabodies (ScDb)wherein two hybrid scFv fragments are connected through a peptide linker(see Kontermann et al (1999) J Immunol Methods 226:179-188). Tetravalentbispecific compounds are available by e.g fusing a scFv fragment to theCH3 domain of an IgG compound or to a Fab fragment through the hingeregion (see Coloma et al (1997) Nature Biotechnol, 15:159-163).Alternatively, tetravalent bispecific compounds have been created by thefusion of bispecific single chain diabodies (see Alt et al (1999) FEBSLett 454:90-94). Smaller tetravalent bispecific compounds can also beformed by the dimerization of either scFv-scFv tandems with a linkercontaining a helix-loop-helix motif (DiBi miniantibodies, see Muller etal (1998) FEBS Lett 432:45-49) or a single chain compound comprisingfour isolated antibody variable domains (VH and VL) in an orientationpreventing intramolecular pairing (tandem diabody, see Kipriyanov et al,(1999) J Mol Biol 293:41-56). Bispecific F(ab′)₂ fragments can becreated by chemical coupling of Fab′ fragments or by heterodimerizationthrough leucine zippers (see Shalaby et al (1992) J Exp Med 175:217-225and Kostelny et al (1992), J Immunol 148:1547-1553).

The term “single variable domain antibody” refers to an antibodyvariable domain (VH, V_(HH), VL) that specifically binds an antigen orepitope independently of a different V region or domain. A singlevariable domain antibody can be present in a format (e.g., homo- orhetero-multimer) with other, different variable regions or variabledomains where the other regions or domains are not required for antigenbinding by the single variable domain (i.e., where the single domainantibody binds antigen independently of the additional variabledomains). A “domain antibody” or “dAb” is the same as a “single variabledomain antibody” which is capable of binding to an antigen as the termis used herein. A single variable domain antibody may be a humanantibody variable domain, but also includes single antibody variabledomains from other species such as rodent (for example, as disclosed inWO 00/29004), nurse shark and camelid V_(HH) dAbs. Camelid V_(HH) aresingle variable domain polypeptides that are derived from speciesincluding camel, llama, alpaca, dromedary, and guanaco, which produceheavy chain antibodies naturally devoid of light chains. Such V_(HH)domains may be humanised according to standard techniques available inthe art, and such domains are still considered to be “domain antibodies”according to the invention. As used herein “VH” includes camelid V_(HH)domains.

2.4) Homologous Antibodies or Fragments Thereof and ConservativeVariants

In another embodiment of the present invention, the binding molecule isan antibody or a fragment thereof which has variable region heavy andlight chain amino acid sequences or heavy and light chain amino acidsequences that are homologous to the amino acid sequences of theantibodies described herein, and wherein the homologous antibodies orfragment thereof retain the desired functional properties of the bindingmolecule according to the invention.

For example, the invention provides an isolated antibody or fragmentthereof comprising a VH and a VL, wherein: the VH is at least 80%, or atleast 90% identical to an amino acid sequence selected from the groupconsisting of SEQ ID NOs:14; 18; 22; 25; 28; 31; 34; 37; 40; 83; 87; 90;93; 112; 130 or 138; the VL is at least 80%, or at least 90% identicalto an amino acid sequence selected from the group consisting of SEQ IDNOs:16; 20; 85; 114; 132; 140; 147 or 153, wherein the homologousantibody specifically binds to IL18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex. The homologous antibody may exhibitat least one additional functional properties such as inhibiting IL18binding to IL18R or inhibiting IL18 dependent IFN-γ production.

In other embodiments, the VH and/or VL amino acid sequences may be 50%,60%, 70%, 80%, 90%, 95%, 96%, 97%, 98% or 99% identical to the sequencesset forth above. In other embodiments, the VH and/or VL amino acidsequences may be identical except an amino acid substitution in no morethan 1, 2, 3, 4 or 5 amino acid positions. An antibody having VH and VLregions having high (i. e., 80% or greater) identity to the VH and VLregions of SEQ ID NOs 14; 18; 22; 25; 28; 31; 34; 37; 40; 83; 87; 90;93; 112; 130 or 138 and SEQ ID NOs 16; 20; 85; 114; 132; 140; 147 or 153respectively, can be obtained by mutagenesis (e.g., site-directed orPCR-mediated mutagenesis) of nucleic acid molecules encoding SEQ ID NOs:15; 19; 23; 26; 29; 32; 35; 38; 41; 84; 88; 91; 94; 113; 131; 139; 146or 152 and 17; 21; 24; 27; 30; 33; 36; 39; 42; 86; 89; 92; 95; 115; 133;141; 148 or 154, respectively, followed by testing of the encodedaltered antibody for retained function (i. e., the functions set forthabove) using the functional assays described herein.

The homologous antibody can be, for example, a human antibody, ahumanized antibody or a chimeric antibody. Preferably the antibody is afully human silent IgG1 antibody.

As used herein, the percent identity between the two sequences is afunction of the number of identical positions shared by the sequences(i. e., % identity=# of identical positions/total # of positions×100),taking into account the number of gaps, and the length of each gap,which need to be introduced for optimal alignment of the two sequences.The comparison of sequences and determination of percent identitybetween two sequences can be accomplished using a mathematicalalgorithm, as described below.

The percent identity between two amino acid sequences can be determinedusing the algorithm of E. Meyers and W. Miller, (1988) Comput. Appl.Biosci 4:11-17 which has been incorporated into the ALIGN program(version 2.0), using a PAM120 weight residue table, a gap length penaltyof 12 and a gap penalty of 4. Alternatively, the percent identitybetween two amino acid sequences can be determined using the Needlemanand Wunsch (1970) J Mol Biol 48:444-453, algorithm which has beenincorporated into the GAP program in the GCG software package (availableat www.gcg.com), using either a Blossom 62 matrix or a PAM250 matrix,and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1,2, 3, 4, 5, or 6.

2.5) Dual Variable Domain Antibodies

Dual variable domain (DVD) antibodies comprise two or more antigenbinding sites and are tetravalent or multivalent antibodies, as forexample divalent and tetravalent. The multivalent antibody isparticularly engineered to have two or more antigen binding sites, andis generally not a naturally occurring antibody. The DVD antibodies maybe capable of binding two or more related or unrelated targets. Such DVDantibodies may be monospecific, i.e capable of binding one antigen ormultispecific, i.e. capable of binding two or more antigens. In someembodiments the DVD antibody comprises two heavy chains and two lightchains. Each heavy chain and light chain comprises two antigen bindingsites. Each binding site comprises a heavy chain variable domain and alight chain variable domain with a total of 6 CDRs involved in antigenbinding per antigen binding site.

In one embodiment, the binding molecule of the present invention is adual variable domain (DVD) antibody which binds IL-18 and does not bindthe IL-18/IL-18 binding protein (IL-18 BP) complex.

Particularly the dual variable domain antibody according to theinvention is capable of binding IL-18 and a second target. The secondtarget can be selected from IL-1, IL-6, IL-8, IL-11, IL-12, IL-17,IL-25, IL-33, IL-1β, TNF alpha/beta and IFN-γ.

2.6) Bispecific and Multispecific Molecules and Antibodies

In one embodiment, the binding molecule of the present invention is anisolated bispecific antibody or fragment thereof comprising a firstspecificity to IL-18 and a second specificity to another polypeptide,e.g. IL-12; wherein the bispecific antibody does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex.

Particularly the bispecific antibody according to the invention iscapable of binding IL-18 and a second target. The second target can beselected from IL-1, IL-6, IL-8, IL-11, IL-12, IL-17, IL-25, IL-33,IL-1β, TNF alpha/beta and IFN-γ.

In yet another aspect, the present invention provides for a bindingmolecule which is an isolated multispecific antibody or fragment thereofcomprising a first specificity to IL-18, a second specificity to anotherpolypeptide, e.g. IL-12 and at least a third specificity to an anotherpolypeptide, wherein the multispecific antibody does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex.

Particularly the multispecific antibody according to the invention iscapable of binding IL-18, a second and a third target. The second andthird targets can be selected from IL-1, IL-6, IL-8, IL-11, IL-12,IL-17, IL-25, IL-33, IL-1β, TNF alpha/beta and IFN-γ.

A bispecific isolated antibody is an isolated antibody having bindingspecificities for at least two different epitopes. Methods of makingsuch antibodies are known in the art. Traditionally, the recombinantproduction of bispecific antibodies is based on the coexpression of twoimmunoglobulin H chain-L chain pairs, where the two H chains havedifferent binding specificities, (see Millstein et al, (1983) Nature305:537-539, WO93/08829 and Traunecker et al, (1991) EMBO 10:3655-3659).Because of the random assortment of H and L chains, a potential mixtureof ten different isolated antibody structures are produced of which onlyone has the desired binding specificity. An alternative approachinvolves fusing the variable domains with the desired bindingspecificities to heavy chain constant region comprising at least part ofthe hinge region, CH2 and CH3 regions. It is preferred to have the CH1region containing the site necessary for light chain binding present inat least one of the fusions. DNA encoding these fusions and, if desired,the L chain, are inserted into separate expression vectors and are thenco-transfected into a suitable host organism. It is possible though toinsert the coding sequences for two or all three chains into oneexpression vector. In one preferred approach, the bispecific isolatedantibody is composed of an H chain with a first binding specificity inone arm and an H-L chain pair, providing a second binding specificity inthe other arm, see WO94/04690. Also see Suresh et al, Methods inEnzymology 121, 210, 1986.

Bispecific and multispecific antibodies of the invention can bederivatized or linked to another functional molecule, e.g., anotherpeptide or protein (e.g., another antibody or ligand for a receptor) togenerate a bispecific molecule that binds to at least two differentbinding sites or target molecules. An antibody of the invention may infact be derivatized or linked to more than one other functional moleculeto generate multi-specific molecules that bind to more than twodifferent binding sites and/or target molecules; such multi-specificmolecules are also intended to be encompassed by the term “bispecificmolecule” as used herein. To create a bispecific molecule of theinvention, an antibody of the invention can be functionally linked(e.g., by chemical coupling, genetic fusion, noncovalent association orotherwise) to one or more other binding molecules, such as anotherantibody, antibody fragment, peptide or binding mimetic, such that abispecific molecule results.

In one embodiment, the bispecific molecules of the invention comprise asa binding specificity at least one antibody, or an antibody fragmentthereof, including, e.g., an Fab, Fab′, F(ab′)₂, Fv, or a single chainFv. The antibody may also be a light chain or heavy chain dimer, or anyminimal fragment thereof such as a Fv or a single chain construct asdescribed in Ladner et al. U.S. Pat. No. 4,946,778.

Other antibodies which can be employed in the bispecific molecules ofthe invention are murine, chimeric and humanized monoclonal antibodies.

The bispecific and multispecific molecules of the present invention canbe prepared by conjugating the constituent binding specificities, usingmethods known in the art. For example, each binding-specificity of thebispecific molecule can be generated separately and then conjugated toone another. When the binding specificities are proteins or peptides, avariety of coupling or cross-linking agents can be used for covalentconjugation. Examples of cross-linking agents include protein A,carbodiimide, N-succinimidyl-S-acetyl-thioacetate (SATA),5,5′-dithiobis(2-nitrobenzoic acid) (DTNB), o-phenylenedimaleimide(oPDM), N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP), andsulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate(sulfo-SMCC) (see e.g., Karpovsky et al (1984) J Exp Med; 160:1686; Liu,M A et al (1985) Proc Natl Acad Sci USA; 82:8648). Other methods includethose described in Paulus (1985) Behring Inst Mitt; 78:118-132; Brennanet al (1985) Science; 229:81-83), and Glennie et al (1987) J Immunol;139:2367-2375. Conjugating agents are SATA and sulfo-SMCC, bothavailable from Pierce Chemical Co. (Rockford, Ill.).

When the binding specificities are antibodies, they can be conjugated bysulphydryl bonding of the C-terminus hinge regions of the two heavychains. In a particular embodiment, the hinge region is modified tocontain an odd number of sulfhydryl residues, for example one, prior toconjugation.

Alternatively, both binding specificities can be encoded in the samevector and expressed and assembled in the same host cell. This method isparticularly useful where the bispecific molecule is a mAb×mAb, mAb×Fab,Fab×F(ab′)₂ or ligand x Fab fusion protein. A bispecific molecule of theinvention can be a single chain molecule comprising one single chainantibody and a binding determinant, or a single chain bispecificmolecule comprising two binding determinants. Bispecific molecules maycomprise at least two single chain molecules. Methods for preparingbispecific molecules are described for example in U.S. Pat. Nos.5,260,203; 5,455,030; 4,881,175; 5,132,405; 5,091,513; 5,476,786;5,013,653; 5,258,498; and 5,482,858.

Binding of the bispecific molecules to their specific targets can beconfirmed by, for example, enzyme-linked immunosorbent assay (ELISA),radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growthinhibition), or Western Blot assay. Each of these assays generallydetects the presence of protein-antibody complexes of particularinterest by employing a labeled reagent (e.g., an antibody) specific forthe complex of interest.

2.7) Multivalent Antibodies

In another aspect, the present invention provides multivalent antibodiescomprising at least two identical or different antigen-binding portionsof the antibodies of the invention binding to IL-18 and not binding theIL-18/IL-18 binding protein (IL-18 BP) complex.

In one embodiment, the multivalent antibody provides at least two, threeor four antigen-binding portions of the antibodies described herein. Theantigen-binding portions can be linked together via protein fusion orcovalent or non covalent linkage. Alternatively, methods of linkage havebeen described for the bispecific molecules. Tetravalent compounds canbe obtained for example by cross-linking antibodies of the inventionwith an antibody that binds to the constant regions of the antibodies ofthe invention, for example the Fc or hinge region.

2.8) Immunoconjugates

In another embodiment, the binding molecule of the present invention isan antibody or a fragment thereof which binds to IL-18 but does not bindthe IL-18/IL-18 binding protein (IL-18 BP) complex, wherein the antibodyor fragment thereof is conjugated to a therapeutic moiety, such as acytotoxin, a drug (e.g., an immunosuppressant) or a radiotoxin.

Such conjugates are referred to herein as “immunoconjugates”.Immunoconjugates that include one or more cytotoxins are referred to as“immunotoxins.” A cytotoxin or cytotoxic agent includes any agent thatis detrimental to (e.g., kills) cells. Examples include taxol,cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin,etoposide, tenoposide, vincristine, vinblastine, t. colchicin,doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone,mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids,procaine, tetracaine, lidocaine, propranolol, and puromycin and analogsor homologs thereof. Therapeutic agents also include, for example,antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine,cytarabine, 5-fluorouracil decarbazine), ablating agents (e.g.,mechlorethamine, thiotepa chlorambucil, meiphalan, carmustine (BSNU) andlomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol,streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP)cisplatin, anthracyclines (e.g., daunorubicin (formerly daunomycin) anddoxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin),bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents(e.g., vincristine and vinblastine).

Other examples of therapeutic cytotoxins that can be conjugated to anantibody of the invention include duocarmycins, calicheamicins,maytansines and auristatins, and derivatives thereof. An example of acalicheamicin antibody conjugate is commercially available (Mylotarg™;Wyeth-Ayerst).

Cytotoxins can be conjugated to antibodies of the invention using linkertechnology available in the art. Examples of linker types that have beenused to conjugate a cytotoxin to an antibody include, but are notlimited to, hydrazones, thioethers, esters, disulfides andpeptide-containing linkers. A linker can be chosen that is, for example,susceptible to cleavage by low pH within the lysosomal compartment orsusceptible to cleavage by proteases, such as proteases preferentiallyexpressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D).

For further discussion of types of cytotoxins, linkers and methods forconjugating therapeutic agents to antibodies, see also Saito, G et al(2003) Adv Drug Deliv Rev; 55:199-215; Trail, P A et al (2003) CancerImmunol Immunother; 52:328-337; Payne, G (2003) Cancer Cell; 3:207 212;Allen, T M (2002) NatRev Cancer; 2:750-763; Pastan, I and Kreitman, R J(2002) Curr Opin Investig Drugs; 3:1089-1091; Senter, P D and Springer,C J (2001) Adv Drug Deliv Rev; 53:247-264.

Antibodies of the present invention also can be conjugated to aradioactive isotope to generate cytotoxic radiopharmaceuticals, alsoreferred to as radioimmunoconjugates. Examples of radioactive isotopesthat can be conjugated to antibodies for use diagnostically ortherapeutically include, but are not limited to, iodine131, indium111,yttrium90, and lutetium177. Methods for preparing radioimmunoconjugatesare established in the art. Examples of radioimmunoconjugates arecommercially available, including Zevalin™ (DEC Pharmaceuticals) andBexxar™ (Corixa Pharmaceuticals), and similar methods can be used toprepare radioimmunoconjugates using the antibodies of the invention.

The antibody conjugates of the invention can be used to modify a givenbiological response, and the drug moiety is not to be construed aslimited to classical chemical therapeutic agents. For example, the drugmoiety may be a protein or polypeptide possessing a desired biologicalactivity. Such proteins may include, for example, an enzymaticallyactive toxin, or active fragment thereof, such as abrin, ricin A,Pseudomonas exotoxin, or diphtheria toxin; or growth factors.

Techniques for conjugating such therapeutic moiety to antibodies arewell known, see, e.g., Amon et al “Monoclonal Antibodies ForImmunotargeting Of Drugs In Cancer Therapy”, in Monoclonal AntibodiesAnd Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss,Inc. 1985); Hellstrom et al “Antibodies For Drug Delivery”, inControlled Drug Delivery (2nd Ed.), Robinson et al (eds.), pp. 623-53(Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of CytotoxicAgents In Cancer Therapy: A Review”, in Monoclonal Antibodies '84:Biological And Clinical Applications, Pinchera et al. (eds.), pp.475-506 (1985); “Analysis, Results, And Future Prospective Of TheTherapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, inMonoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al.(eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., “ThePreparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”,(1982) Immunol Rev; 62:119-58.

2.9) Heteroconjugate Antibodies

Heteroconjugate antibodies also form an embodiment of the presentinvention. Heteroconjugate antibodies are composed of two covalentlyjoined antibodies formed using any convenient cross-linking methods,wherein at least of the joined antibodies is an antibody according tothe invention which binds IL-18 but does not bind the IL-18/IL-18binding protein (IL-18 BP) complex. See, for example, U.S. Pat. No.4,676,980.

2.10) Framework Engineering

Engineered antibodies of the invention include those in whichmodifications have been made to framework residues within VH and/or VL,e.g. to improve the properties of the antibody. Typically such frameworkmodifications are made to decrease the immunogenicity of the antibody.For example, one approach is to “backmutate” one or more frameworkresidues to the corresponding germline sequence. More specifically, anantibody that has undergone somatic mutation may contain frameworkresidues that differ from the germline sequence from which the antibodyis derived. Such residues can be identified by comparing the antibodyframework sequences to the germline sequences from which the antibody isderived. To return the framework region sequences to their germlineconfiguration, the somatic mutations can be “backmutated” to thegermline sequence by, for example, site-directed mutagenesis orPCR-mediated mutagenesis. Such “backmutated” antibodies are alsointended to be encompassed by the invention.

Another type of framework modification involves mutating one or moreresidues within the framework region, or even within one or more CDRregions, to remove T cell-epitopes to thereby reduce the potentialimmunogenicity of the antibody. This approach is also referred to as“deimmunization” and is described in further detail in U.S. PatentPublication No. 20030153043 by Carr et al.

2.11) Other Modifications

In addition or alternative to modifications made within the framework orCDR regions, antibodies of the invention may be engineered to includemodifications within the Fc region, typically to alter one or morefunctional properties of the antibody, such as serum half-life,complement fixation, Fc receptor binding, and/or antigen-dependentcellular cytotoxicity. Furthermore, an antibody of the invention may bechemically modified (e.g., one or more chemical moieties can be attachedto the antibody) or be modified to alter its glycosylation, again toalter one or more functional properties of the antibody. Each of theseembodiments is described in further detail below. The numbering ofresidues in the Fc region is that of the EU index of Kabat.

In one embodiment, the hinge region of CH1 is modified such that thenumber of cysteine residues in the hinge region is altered, e.g.,increased or decreased. This approach is described further in U.S. Pat.No. 5,677,425 by Bodmer et al. The number of cysteine residues in thehinge region of CH1 is altered to, for example, facilitate assembly ofthe light and heavy chains or to increase or decrease the stability ofthe antibody.

In another embodiment, the Fc hinge region of an antibody is mutated todecrease the biological half-life of the antibody. More specifically,one or more amino acid mutations are introduced into the CH2-CH3 domaininterface region of the Fc-hinge fragment such that the antibody hasimpaired Staphylococcal protein A (SpA) binding relative to nativeFc-hinge domain SpA binding. This approach is described in furtherdetail in U.S. Pat. No. 6,165,745 by Ward et al. In another embodiment,the antibody is modified to increase its biological half-life. Variousapproaches are possible. For example, one or more of the followingmutations can be introduced: T252L, T254S, T256F, as described in U.S.Pat. No. 6,277,375 to Ward. Alternatively, to increase the biologicalhalf life, the antibody can be altered within the CH1 or CL region tocontain a salvage receptor binding epitope taken from two loops of a CH2domain of an Fc region of an IgG, as described in U.S. Pat. Nos.5,869,046 and 6,121,022 by Presta et al.

In yet other embodiments, the Fc region is altered by replacing at leastone amino acid residue with a different amino acid residue to alter theeffector functions of the antibody. For example, one or more amino acidscan be replaced with a different amino acid residue such that theantibody has an altered affinity for an effector ligand but retains theantigen-binding ability of the parent antibody. The effector ligand towhich affinity is altered can be, for example, an Fc receptor or the C1component of complement. This approach is described in further detail inU.S. Pat. Nos. 5,624,821 and 5,648,260, both by Winter et al.

In another embodiment, one or more amino acids selected from amino acidresidues can be replaced with a different amino acid residue such thatthe antibody has altered C1 q binding and/or reduced or abolishedcomplement dependent cytotoxicity (CDC). This approach is described infurther detail in U.S. Pat. No. 6,194,551 by Idusogie et al.

In another embodiment, one or more amino acid residues are altered tothereby alter the ability of the antibody to fix complement. Thisapproach is described further in PCT Publication WO 94/29351 by Bodmeret al.

In yet another embodiment, the Fc region is modified to increase theability of the antibody to mediate antibody dependent cellularcytotoxicity (ADCC) and/or to increase the affinity of the antibody foran Fcγ receptor by modifying one or more amino acids. This approach isdescribed further in PCT Publication WO 00/42072 by Presta. Moreover,the binding sites on human IgG1 for FcγRI, FcγRII, FcγRIII and FcRn havebeen mapped and variants with improved binding have been described (seeShields, R. L. et al, (2001) J Biol Chem 276:6591-6604).

In certain embodiments, the Fc domain of IgG1 isotype is used. In somespecific embodiments, a mutant variant of IgG1 Fc fragment is used, e.g.a silent IgG1 Fc which reduces or eliminates the ability of the fusionpolypeptide to mediate antibody dependent cellular cytotoxicity (ADCC)and/or to bind to an Fcγ receptor. An example of an IgG1 isotype silentmutant wherein Leucine residue is replaced by Alanine residue at aminoacid positions 234 and 235 as described by Hezareh et al, J. Virol(2001); 75(24):12161-8.

In certain embodiments, the Fc domain is a mutant preventingglycosylation at position 297 of Fc domain. For example, the Fc domaincontains an amino acid substitution of asparagine residue at position297. Example of such amino acid substitution is the replacement of N297by a glycine or an alanine.

In still another embodiment, the glycosylation of an antibody ismodified. For example, an aglycosylated antibody can be made (i.e., theantibody lacks glycosylation). Glycosylation can be altered to, forexample, increase the affinity of the antibody for the antigen. Suchcarbohydrate modifications can be accomplished by; for example, alteringone or more sites of glycosylation within the antibody sequence. Forexample, one or more amino acid substitutions can be made that result inelimination of one or more variable region framework glycosylation sitesto thereby eliminate glycosylation at that site. Such aglycosylation mayincrease the affinity of the antibody for antigen. Such an approach isdescribed in further detail in U.S. Pat. Nos. 5,714,350 and 6,350,861 byCo et al.

Additionally or alternatively, an antibody can be made that has analtered type of glycosylation, such as a hypofucosylated antibody havingreduced amounts of fucosyl residues or an antibody having increasedbisecting GlcNac structures. Such altered glycosylation patterns havebeen demonstrated to increase the ADCC ability of antibodies. Suchcarbohydrate modifications can be accomplished by, for example,expressing the antibody in a host cell with altered glycosylationmachinery. Cells with altered glycosylation machinery have beendescribed in the art and can be used as host cells in which to expressrecombinant antibodies of the invention to thereby produce an antibodywith altered glycosylation. For example, EP 1,176,195 by Hang et al.describes a cell line with a functionally disrupted FUT8 gene, whichencodes a fucosyl transferase, such that antibodies expressed in such acell line exhibit hypofucosylation. Therefore, in one embodiment, theantibodies of the invention are produced by recombinant expression in acell line which exhibit hypofucosylation pattern, for example, amammalian cell line with deficient expression of the FUT8 gene encodingfucosyltransferase. PCT Publication WO 03/035835 by Presta describes avariant CHO cell line, Lecl3 cells, with reduced ability to attachfucose to Asn(297)-linked carbohydrates, also resulting inhypofucosylation of antibodies expressed in that host cell (see alsoShields, R. L. et al., 2002 J. Biol. Chem. 277:26733-26740). PCTPublication WO 99/54342 by Umana et al. describes cell lines engineeredto express glycoprotein-modifying glycosyl transferases (e.g.,beta(1,4)-N acetylglucosaminyltransferase III (GnTIII)) such thatantibodies expressed in the engineered cell lines exhibit increasedbisecting GlcNac structures which results in increased ADCC activity ofthe antibodies (see also Umana et al., 1999 Nat. Biotech. 17:176-180).Alternatively, the antibodies of the invention can be produced in ayeast or a filamentous fungi engineered for mammalian-like glycosylationpattern, and capable of producing antibodies lacking fucose asglycosylation pattern (see for example EP1297172B1).

Another modification of the antibodies herein that is contemplated bythe invention is pegylation. An antibody can be pegylated to, forexample, increase the biological (e.g., serum) half-life of theantibody. To pegylate an antibody, the antibody, or fragment thereof,typically is reacted with polyethylene glycol (PEG), such as a reactiveester or aldehyde derivative of PEG, under conditions in which one ormore PEG groups become attached to the antibody or antibody fragment.The pegylation can be carried out by an acylation reaction or analkylation reaction with a reactive PEG molecule (or an analogousreactive water-soluble polymer). As used herein, the term “polyethyleneglycol” is intended to encompass any of the forms of PEG that have beenused to derivatize other proteins, such as mono (C1-C10) alkoxy- oraryloxy-polyethylene glycol or polyethylene glycol-maleimide. In certainembodiments, the antibody to be pegylated is an aglycosylated antibody.Methods for pegylating proteins are known in the art and can be appliedto the antibodies of the invention. See for example, EP 0 154 316 byNishimura et al. and EP 0 401 384 by Ishikawa et al.

Another modification of the antibodies that is contemplated by theinvention is a conjugate or a protein fusion of at least theantigen-binding region of the antibody of the invention to serumprotein, such as human serum albumin or a fragment thereof to increasehalf-life of the resulting molecule. Such approach is for exampledescribed in Ballance et al. EP0322094.

3. Grafting Antigen-Binding Domains into Alternative Frameworks orScaffolds

A wide variety of antibody/immunoglobulin frameworks or scaffolds can beemployed so long as the resulting polypeptide includes at least onebinding region which specifically binds to IL18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex and wherein the bindingmolecule is not IL-18BP. Such frameworks or scaffolds include the 5 mainidiotypes of human immunoglobulins, or fragments thereof (such as thosedisclosed elsewhere herein), and include immunoglobulins of other animalspecies, preferably having humanized aspects. Single heavy-chainantibodies such as those identified in camelids are of particularinterest in this regard. Novel frameworks, scaffolds and fragmentscontinue to be discovered and developed by those skilled in the art.

3.1) Non-Immunoglobulin Frameworks

Known or future non-immunoglobulin frameworks and scaffolds may beemployed, as long as they comprise a binding region specific for IL-18and not binding the IL-18/IL-18 binding protein (IL-18 BP) complex andwherein the binding molecule is not IL-18BP. Such compounds are referredherein as “polypeptides comprising a target-specific binding region”.Known non-immunoglobulin frameworks or scaffolds include, but are notlimited to, adnectins (Adnectins®-Compound Therapeutics, Inc., Waltham,Mass.), DARPins, avimers, Affibodys® (Affibody AG, Sweden), anticalins(Pieris Proteolab AG, Freising, Germany), Affilins® (gamma-crystallin orubiquitin; Scil Proteins GmbH, Halle, Germany) and protein epitopemimetics (PEM; PolyphorO Ltd, Allschwil, Switzerland).

3.2) Fibronectin Molecules and Adnectins

In one aspect of the invention the binding molecule is a fibronectinmolecule. The fibronectin molecule has a scaffold based preferably onfibronectin type III domain (e.g., the tenth module of the fibronectintype III (10 Fn3 domain)). In one embodiment the binding molecule is anadnectin (Adnectins®).

The fibronectin type III domain has 7 or 8 beta strands which aredistributed between two beta sheets, which themselves pack against eachother to form the core of the protein, and further containing loops(analogous to CDRs) which connect the beta strands to each other and aresolvent exposed. There are at least three such loops at each edge of thebeta sheet sandwich, where the edge is the boundary of the proteinperpendicular to the direction of the beta strands (U.S. Pat. No.6,818,418).

These fibronectin-based scaffolds are not an immunoglobulin, althoughthe overall fold is closely related to that of the smallest functionalantibody fragment, the variable region of the heavy chain, whichcomprises the entire antigen recognition unit in camel and llama IgG.Because of this structure, the non-immunoglobulin antibody mimicsantigen binding properties that are similar in nature and affinity tothose of antibodies. These scaffolds can be used in a loop randomizationand shuffling strategy in vitro that is similar to the process ofaffinity maturation of antibodies in vivo. These fibronectin-basedmolecules can be used as scaffolds where the loop regions of themolecule can be replaced with CDRs of the invention using standardcloning techniques.

Accordingly, in one embodiment the binding molecule is a fibronectinmolecule that binds (e.g. specifically binds) to IL18 and does not bindthe IL-18/IL-18 binding protein (IL-18 BP) complex. In anotherembodiments the binding molecule is an adnectin that binds (e.g.specifically binds) to IL18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex.

3.3) DARPins

The technology is based on using proteins with ankyrin derived repeatmodules as scaffolds for bearing variable regions which can be used forbinding to different targets. The ankyrin repeat module is a 33 aminoacid polypeptide consisting of two anti-parallel α-helices and a β-turnBinding of the variable regions is mostly optimized by using ribosomedisplay.

Accordingly, in one embodiment the binding molecule is anankyrin/DARPins that binds (e.g. specifically binds) to IL18 and doesnot bind the IL-18/IL-18 binding protein (IL-18 BP) complex.

3.4) Avimers

Avimers are derived from natural A-domain containing protein such asLRP-1. These domains are used by nature for protein-protein interactionsand in human over 250 proteins are structurally based on A-domains.Avimers consist of a number of different “A-domain” monomers (2-10)linked via amino acid linkers. Avimers can be created that bind to thetarget antigen using the methodology described in, for example,US20040175756; US20050053973; US20050048512; and US20060008844.

Accordingly, in one embodiment the binding molecule is an avimer thatbinds (e.g. specifically binds) to IL18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex.

3.5) Affibody®

Affibody® are small, simple proteins composed of a three-helix bundlebased on the scaffold of one of the IgG-binding domains of Protein A.Protein A is a surface protein from the bacterium Staphylococcus aureus.This scaffold domain consists of 58 amino acids, 13 of which arerandomized to generate Affibody® libraries with a large number of ligandvariants (See e.g., U.S. Pat. No. 5,831,012). Affibody® molecules mimicantibodies; they have a molecular weight of 6 kDa, compared to themolecular weight of antibodies, which is 150 kDa. In spite of its smallsize, the binding site of Affibody® molecules is similar to that of anantibody.

Accordingly, in one embodiment the binding molecule is an affibody thatbinds (e.g. specifically binds) to IL18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex.

3.6) Anticalins®

Anticalins® are products developed by the company Pieris ProteoLab AG.They are derived from lipocalins, a widespread group of small and robustproteins that are usually involved in the physiological transport orstorage of chemically sensitive or insoluble compounds. Several naturallipocalins occur in human tissues or body liquids.

The protein architecture is reminiscent of immunoglobulins, withhypervariable loops on top of a rigid framework. However, in contrastwith antibodies or their recombinant fragments, lipocalins are composedof a single polypeptide chain with 160 to 180 amino acid residues, beingjust marginally bigger than a single immunoglobulin domain.

The set of four loops, which makes up the binding pocket, showspronounced structural plasticity and tolerates a variety of side chains.The binding site can thus be reshaped in a proprietary process in orderto recognize prescribed target compounds of different shape with highaffinity and specificity.

One protein of the lipocalin family, the bilin-binding protein (BBP) ofPieris Brassicae has been used to develop anticalins by mutagenizing theset of four loops. One example of a patent application describing“anticalins” is PCT WO 199916873.

Accordingly, in one embodiment the binding molecule is an anticalin thatbinds (e.g. specifically binds) to IL18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex.

3.7) Affilin®

Affilin® molecules are small non-immunoglobulin proteins which aredesigned for specific affinities towards proteins and small molecules.New Affilin® molecules can be very quickly selected from two libraries,each of which is based on a different human derived scaffold protein.

Affilin™ molecules do not show any structural homology to immunoglobulinproteins. Scil Proteins employs two Affilin™ scaffolds, one of which isgamma crystalline, a human structural eye lens protein and the other is“ubiquitin” superfamily proteins. Both human scaffolds are very small,show high temperature stability and are almost resistant to pH changesand denaturing agents. This high stability is mainly due to the expandedbeta sheet structure of the proteins. Examples of gamma crystallinederived proteins are described in WO200104144 and examples of“ubiquitin-like” proteins are described in WO2004106368 Accordingly, inone embodiment the binding molecule is an affilin that binds (e.g.specifically binds) to IL18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex.

3.8) Protein Epitope Mimetics

Protein Epitope Mimetics (PEM) are medium-sized, cyclic, peptide-likemolecules (ca. 1-2 kDa) mimicking beta-hairpin secondary structures ofproteins, the major secondary structure involved in protein-proteininteractions.

Accordingly, in one embodiment the binding molecule is a protein epitopemimetic that binds (e.g. specifically binds) to IL18 and does not bindthe IL-18/IL-18 binding protein (IL-18 BP) complex.

4. Polynucleotides Encoding the Binding Molecules of the Invention

Another aspect of the invention pertains to isolated polynucleotidesencoding the binding molecules of the invention.

Heavy chain variable domain polynucleotides encoding the isolated humanantibodies described herein are shown in SEQ ID NOs: 15, 19, 23, 26, 29,32, 35, 38, 41, 84, 88, 91, 94, 113, 131, 139, 146 and 152. Light chainvariable domain polynucleotides encoding the isolated human antibodiesdescribed herein are shown in SEQ ID NOs: 17, 21, 24, 27, 30, 33, 36,39, 42, 86, 89, 92, 95, 115, 133, 141, 148 and 154. Otherpolynucleotides encoding antibodies of the invention includepolynucleotides that have been mutated, yet have at least 60, 70, 80, 90or 95 percent identity to the sequences described above such aspolynucleotides which have been optimized for protein expression inmammalian cells, for example, CHO cell lines.

Embodiment 89: A variant nucleic acids wherein no more than 1, 2, 3, 4or 5 nucleotides have been changed by nucleotide deletion, insertion orsubstitution in the variable regions when compared with the variableregions depicted in the sequences described above.

Embodiment 90: The isolated polynucleotide which encodes the heavy chainvariable domain of an antibody or fragment thereof according to theinvention, wherein the polynucleotide:

a. is at least 90% identical to SEQ ID NO: 15 or SEQ ID NO: 19 or SEQ IDNO: 23 or SEQ ID NO: 26 or SEQ ID NO: 29 or SEQ ID NO: 32 or SEQ ID NO:35 or SEQ ID NO: 38 or SEQ ID NO: 41 or SEQ ID NO: 84 or SEQ ID NO: 88or SEQ ID NO: 91 or SEQ ID NO: 94 or SEQ ID NO: 113 or SEQ ID NO: 131 orSEQ ID NO: 139 or SEQ ID NO: 146 or SEQ ID NO: 152 or

b. comprises SEQ ID NO: 15 or SEQ ID NO: 19 or SEQ ID NO: 23 or SEQ IDNO: 26 or SEQ ID NO: 29 or SEQ ID NO: 32 or SEQ ID NO: 35 or SEQ ID NO:38 or SEQ ID NO: 41 or SEQ ID NO: 84 or SEQ ID NO: 88 or SEQ ID NO: 91or SEQ ID NO: 94 or SEQ ID NO: 113 or SEQ ID NO: 131 or SEQ ID NO: 139or SEQ ID NO: 146 or SEQ ID NO: 152 or

c. consists essentially of SEQ ID NO: 15 or SEQ ID NO: 19 or SEQ ID NO:23 or SEQ ID NO: 26 or SEQ ID NO: 29 or SEQ ID NO: 32 or SEQ ID NO: 35or SEQ ID NO: 38 or SEQ ID NO: 41 or SEQ ID NO: 84 or SEQ ID NO: 88 orSEQ ID NO: 91 or SEQ ID NO: 94 or SEQ ID NO: 113 or SEQ ID NO: 131 orSEQ ID NO: 139 or SEQ ID NO: 146 or SEQ ID NO: 152.

Embodiment 91: The isolated polynucleotide which encodes the light chainvariable domain of an antibody or a fragment thereof according to theinvention, wherein the polynucleotide:

a. is at least 90% identical to SEQ ID NO: 17 or SEQ ID NO: 21 or SEQ IDNO: 24 or SEQ ID NO: 27 or SEQ ID NO: 30 or SEQ ID NO: 33 or SEQ ID NO:36 or SEQ ID NO: 39 or SEQ ID NO: 42 or SEQ ID NO: 86 or SEQ ID NO: 89or SEQ ID NO: 92 or SEQ ID NO: 95 or SEQ ID NO: 115 or SEQ ID NO: 133 orSEQ ID NO: 141 or SEQ ID NO: 148 or SEQ ID NO: 154 or

b. comprises SEQ ID NO: 17 or SEQ ID NO: 21 or SEQ ID NO: 24 or SEQ IDNO: 27 or SEQ ID NO: 30 or SEQ ID NO: 33 or SEQ ID NO: 36 or SEQ ID NO:39 or SEQ ID NO: 42 or SEQ ID NO: 86 or SEQ ID NO: 89 or SEQ ID NO: 92or SEQ ID NO: 95 or SEQ ID NO: 115 or SEQ ID NO: 133 or SEQ ID NO: 141or SEQ ID NO: 148 or SEQ ID NO: 154 or

c. consists essentially of SEQ ID NO: 17 or SEQ ID NO: 21 or SEQ ID NO:24 or SEQ ID NO: 27 or SEQ ID NO: 30 or SEQ ID NO: 33 or SEQ ID NO: 36or SEQ ID NO: 39 or SEQ ID NO: 42 or SEQ ID NO: 86 or SEQ ID NO: 89 orSEQ ID NO: 92 or SEQ ID NO: 95 or SEQ ID NO: 115 or SEQ ID NO: 133 orSEQ ID NO: 141 or SEQ ID NO: 148 or SEQ ID NO: 154.

Embodiment 92: The isolated polynucleotide which encodes the heavy chainof an antibody according to the invention, wherein the polynucleotide:

a. is at least 90% identical to SEQ ID NO: 44 or SEQ ID NO: 48 or SEQ IDNO: 51 or SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO: 101 or SEQ ID NO:159 or SEQ ID NO: 97 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID NO:143 or SEQ ID NO: 135 or SEQ ID NO: 149 or SEQ ID NO: 155; or

b. comprises SEQ ID NO: 44 or SEQ ID NO: 48 or SEQ ID NO: 51 or SEQ IDNO: 54 or SEQ ID NO: 57 or SEQ ID NO: 101 or SEQ ID NO: 159 or SEQ IDNO: 97 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID NO: 143 or SEQ IDNO: 135 or SEQ ID NO: 149 or SEQ ID NO: 155; or

c. consists essentially of SEQ ID NO: 44 or SEQ ID NO: 48 or SEQ ID NO:51 or SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO: 101 or SEQ ID NO: 159or SEQ ID NO: 97 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ ID NO: 143or SEQ ID NO: 135 or SEQ ID NO: 149 or SEQ ID NO: 155.

Embodiment 93: The isolated polynucleotide encodes the light chain of anantibody according to the invention, wherein the polynucleotide:

a. is at least 90% identical to SEQ ID NO: 46 or SEQ ID NO: 49 or SEQ IDNO: 52 or SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO: 102 or SEQ ID NO:161 or SEQ ID NO: 99 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ ID NO:145 or SEQ ID NO: 137 or SEQ ID NO: 151 or SEQ ID NO: 157; or

b. comprises SEQ ID NO: 46 or SEQ ID NO: 49 or SEQ ID NO: 52 or SEQ IDNO: 55 or SEQ ID NO: 58 or SEQ ID NO: 102 or SEQ ID NO: 161 or SEQ IDNO: 99 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ ID NO: 145 or SEQ IDNO: 137 or SEQ ID NO: 151 or SEQ ID NO: 157; or

c. consists essentially of SEQ ID NO: 46 or SEQ ID NO: 49 or SEQ ID NO:52 or SEQ ID NO: 55 or SEQ ID NO: 58 or SEQ ID NO: 102 or SEQ ID NO: 161or SEQ ID NO: 99 or SEQ ID NO: 105 or SEQ ID NO: 119 or SEQ ID NO: 145or SEQ ID NO: 137 or SEQ ID NO: 151 or SEQ ID NO: 157.

The polynucleotide may be present in whole cells, in a cell lysate, ormay be in a partially purified or substantially pure form. Apolynucleotide is “isolated” or “rendered substantially pure” whenpurified away from other cellular components or other contaminants,e.g., other cellular nucleic acids or proteins, by standard techniques,including alkaline/SDS treatment, CsCl banding, column chromatography,agarose gel electrophoresis and others well known in the art. See, F.Ausubel, et al., (1987) Current Protocols in Molecular Biology, GreenePublishing and Wiley Interscience, New York. A polynucleotide of theinvention can be, for example, DNA or RNA and may or may not containintronic sequences. In an embodiment, the polynucleotide is a cDNAmolecule. The polynucelotide may be present in a vector such as a phagedisplay vector, or in a recombinant plasmid vector.

Polynucleotides of the invention can be obtained using standardmolecular biology techniques. For antibodies expressed by hybridomas(e.g., hybridomas prepared from transgenic mice carrying humanimmunoglobulin genes as described further below), cDNAs encoding thelight and heavy chains of the antibody made by the hybridoma can beobtained by standard PCR amplification or cDNA cloning techniques. Forantibodies obtained from an immunoglobulin gene library (e.g., usingphage display techniques), polynucleotides encoding the antibody can berecovered from various phage clones that are members of the library.

Once DNA fragments encoding VH and VL segments are obtained, these DNAfragments can be further manipulated by standard recombinant DNAtechniques, for example to convert the variable region genes tofull-length antibody chain genes, to Fab fragment genes or to an scFvgene. In these manipulations, a VL- or VH-encoding DNA fragment isoperatively linked to another DNA molecule, or to a fragment encodinganother protein, such as an antibody constant region or a flexiblelinker. The term “operatively linked”, as used in this context, isintended to mean that the two DNA fragments are joined in a functionalmanner, for example, such that the amino acid sequences encoded by thetwo DNA fragments remain in-frame, or such that the protein is expressedunder control of a desired promoter.

The isolated DNA encoding the VH region can be converted to afull-length heavy chain gene by operatively linking the VH-encoding DNAto another DNA molecule encoding heavy chain constant regions (CH1, CH2and CH3). The sequences of human heavy chain constant region genes areknown in the art (see e.g., Kabat, E. A., el al., (1991) Sequences ofProteins of Immunological Interest, Fifth Edition, U.S. Department ofHealth and Human Services, NIH Publication No. 91-3242) and DNAfragments encompassing these regions can be obtained by standard PCRamplification. The heavy chain constant region can be an IgG1, IgG2,IgG3, IgG4, IgA, IgE, IgM or IgD constant region. In some embodiments,the heavy chain contstant region is selected among IgG1 isotypes. For aFab fragment heavy chain gene, the VH-encoding DNA can be operativelylinked to another DNA molecule encoding only the heavy chain CH1constant region.

The isolated DNA encoding the VL region can be converted to afull-length light chain gene (as well as to a Fab light chain gene) byoperatively linking the VL-encoding DNA to another DNA molecule encodingthe light chain constant region, CL. The sequences of human light chainconstant region genes are known in the art (see e.g., Kabat, E. A., etal., (1991) Sequences of Proteins of Immunological Interest, FifthEdition, U.S. Department of Health and Human Services, NIH PublicationNo. 91-3242) and DNA fragments encompassing these regions can beobtained by standard PCR amplification. The light chain constant regioncan be a kappa or a lambda constant region.

To create an scFv gene, the VH- and VL-encoding DNA fragments areoperatively linked to another fragment encoding a flexible linker, e.g.,encoding the amino acid sequence (Gly4-Ser)3, such that the VH and VLsequences can be expressed as a contiguous single-chain protein, withthe VL and VH regions joined by the flexible linker (see e.g., Bird etal., 1988 Science 242:423-426; Huston et at., (1988) Proc. Natl. Acad.Sci. USA 85:5879-5883; McCafferty et al., (1990) Nature 348:552-554).

The polynucleotide according to any one of embodiments 89 to 93 may bein-vitro modified in such a way that when injected into a mammaliancells, it prevents digestion of the polynucleotide and allows thetranslation machinery of the mammalian cell to produce an antibodystarting from the modified polynucleotide (such as described in U.S.Pat. No. 8,278,036 B2 to Kariko et al.). Such an in vitro-synthesizedmodified RNA can then be injected into a mammalian cells or into apatient as part of a gene therapy.

Hence, another aspect of the present invention encompasses a method forinducing a mammalian cell to produce an antibody as described herein,the method comprising: contacting said mammalian cell with invitro-synthesized modified RNA derived from a polynucleotide accordingto any one of embodiments 89 to 93, wherein said in vitro-synthesizedmodified RNA comprises one or more modification as described in U.S.Pat. No. 8,278,036 B2.

5. Production of Antibodies

Monoclonal antibodies (mAbs) can be produced by a variety of techniques,including conventional monoclonal antibody methodology e.g., thestandard somatic cell hybridization technique of Kohler and Milstein,(1975) Nature 256: 495. Many techniques for producing monoclonalantibody can be employed e.g., viral or oncogenic transformation of Blymphocytes.

Immunization protocols and techniques for isolation of immunizedsplenocytes for fusion are known in the art. Fusion partners (e.g.,murine myeloma cells) and fusion procedures are also known.

Chimeric or humanized monoclonal antibodies of the present invention canbe prepared based on the sequence of a murine monoclonal antibodyprepared as described above. DNA encoding the heavy and light chain ofimmunoglobulins can be obtained from the murine hybridoma of interestand engineered to contain non-murine (e.g., human) immunoglobulinsequences using standard molecular biology techniques. For example, tocreate a chimeric antibody, the murine variable regions can be linked tohuman constant regions using methods known in the art (see e.g., U.S.Pat. No. 4,816,567 to Cabilly et al.). To create a humanized antibody,the murine CDR regions can be inserted into a human framework usingmethods known in the art. See e.g., U.S. Pat. No. 5,225,539 to Winter,and U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,762 and 6,180,370 toQueen et al.

Human monoclonal antibodies can be generated using transgenic ortranschromosomic mice carrying parts of the human immune system ratherthan the mouse system. These transgenic and transchromosomic miceinclude mice referred to herein as HuMAb mice and KM mice, respectively,and are collectively referred to herein as “human Ig mice.”

The HuMAb Mouse® (Medarex, Inc.) contains human immunoglobulin geneminiloci that encode un-rearranged human heavy (μ and γ) and K lightchain immunoglobulin sequences, together with targeted mutations thatinactivate the endogenous p and K chain loci (see e.g., Lonberg et al(1994) Nature; 368(6474): 856-859). Accordingly, the mice exhibitreduced expression of mouse IgM or κ, and in response to immunization,the introduced human heavy and light chain transgenes undergo classswitching and somatic mutation to generate high affinity human IgGκmonoclonal (Lonberg, N et al (1994) supra; reviewed in Lonberg, N (1994)Handbook of Experimental Pharmacology 113:49-101; Lonberg, N and Huszar,D (1995) Intern Rev Immunol; 13: 65-93, and Harding, F and Lonberg, N(1995) Ann N Y Acad Sci; 764:536 546). The preparation and use of HuMAbmice, and the genomic modifications carried by such mice, is furtherdescribed in Taylor, L et al; (1992) Nucl Acids Res; 20:6287-6295; Chen,J et al (1993) Int Immunol; 5: 647-656; Tuaillon et al (1993) Proc NatlAcad Sci USA; 94:3720-3724; Choi et al (1993) Nature Gen; 4:117-123;Chen, J et al (1993) EMBO J; 12: 821-830; Tuaillon et al (1994) JImmunol; 152:2912-2920; Taylor, L et al (1994) Int Immuno; 6:579-591;and Fishwild, D et al (1996) Nature Biotech; 14: 845-851, the contentsof all of which are hereby specifically incorporated by reference intheir entirety. See further, U.S. Pat. Nos. 5,545,806; 5,569,825;5,625,126; 5,633,425; 5,789,650; 5,877,397; 5,661,016; 5,814,318;5,874,299; and 5,770,429; all to Lonberg and Kay; U.S. Pat. No.5,545,807 to Surani et al; PCT Publication Nos. WO 92103918, WO93/12227, WO 94/25585, WO 97113852, WO 98/24884 and WO 99/45962, all toLonberg and Kay; and PCT Publication No. WO 01/14424 to Korman et al.

In one embodiment, human antibodies according to the invention can beraised using a mouse that carries human immunoglobulin sequences ontransgenes and transchomosomes such as a mouse that carries a humanheavy chain transgene and a human light chain transchromosome. Suchmice, referred to herein as “KM mice”, are described in detail in PCTPublication WO 02/43478 to Ishida et al.

Still further, alternative transgenic animal systems expressing humanimmunoglobulin genes are available in the art and can be used to raiseantibodies of the invention. For example, an alternative transgenicsystem referred to as the Xenomouse (Abgenix, Inc.) can be used. Suchmice are described in, e.g., U.S. Pat. Nos. 5,939,598; 6,075,181;6,114,598; 6, 150,584 and 6,162,963 to Kucherlapati et al.

Moreover, alternative transchromosomic animal systems expressing humanimmunoglobulin genes are available in the art and can be used to raiseantibodies of the invention. For example, mice carrying both a humanheavy chain transchromosome and a human light chain tranchromosome,referred to as “TC mice” can be used; such mice are described inTomizuka et al., 2000 Proc. Natl. Acad. Sci. USA 97:722-727.Furthermore, cows carrying human heavy and light chain transchromosomeshave been described in the art (Kuroiwa et al., 2002 Nature Biotech20:889-894).

Human recombinant antibodies of the invention can also be prepared usingphage display methods for screening libraries of human immunoglobulingenes. Such phage display methods for isolating human antibodies areestablished in the art or described in the examples below. See forexample: U.S. Pat. Nos. 5,223,409; 5,403,484; and U.S. Pat. No.5,571,698 to Ladner et al.; U.S. Pat. Nos. 5,427,908 and 5,580,717 toDower et al.; U.S. Pat. Nos. 5,969,108 and 6,172,197 to McCafferty etal.; and U.S. Pat. Nos. 5,885,793; 6,521,404; 6,544,731; 6,555,313;6,582,915 and 6,593,081 to Griffiths et al.

Human monoclonal antibodies of the invention can also be prepared usingSLID mice into which human immune cells have been reconstituted suchthat a human antibody response can be generated upon immunization. Suchmice are described in, for example, U.S. Pat. Nos. 5,476,996 and5,698,767 to Wilson et al.

Antibodies of the invention also can be produced in a host celltransfectoma using, for example, a combination of recombinant DNAtechniques and gene transfection methods as is well known in the art(e.g., Morrison, S. (1985) Science 229:1202).

For example, to express the antibodies, or antibody fragments thereof,DNAs encoding partial or full-length light and heavy chains, can beobtained by standard molecular biology techniques (e.g., PCRamplification or cDNA cloning using a hybridoma that expresses theantibody of interest) and the DNAs can be inserted into cloning orexpression vectors such that the genes are operatively linked totranscriptional and translational control sequences. In this context,the term “operatively linked” is intended to mean that an antibody geneis ligated into a vector such that transcriptional and translationalcontrol sequences within the vector serve their intended function ofregulating the transcription and translation of the antibody gene. Thecloning or expression vector and expression control sequences are chosento be compatible with the expression host cell used. The antibody lightchain gene and the antibody heavy chain gene can be inserted intoseparate vector or, more typically, both genes are inserted into thesame expression vector. The antibody genes are inserted into theexpression vector by standard methods (e.g., ligation of complementaryrestriction sites on the antibody gene fragment and vector, or blunt endligation if no restriction sites are present). The light and heavy chainvariable regions of the antibodies described herein can be used tocreate full-length antibody genes of any antibody isotype by insertingthem into expression vectors already encoding heavy chain constant andlight chain constant regions of the desired isotype such that the VHsegment is operatively linked to the CH segment(s) within the vector andthe VL segment is operatively linked to the CL segment within thevector. Additionally or alternatively, the recombinant expression vectorcan encode a signal peptide that facilitates secretion of the antibodychain from a host cell. The antibody chain gene can be cloned into thevector such that the signal peptide is linked in frame to the aminoterminus of the antibody chain gene. The signal peptide can be animmunoglobulin signal peptide or a heterologous signal peptide (i.e., asignal peptide from a non-immunoglobulin protein).

In one aspect, the invention provides a cloning or expression vectorcomprising one or more polynucleotides according to the invention.

Embodiment 94: The cloning or expression vector which comprises at leastone polynucleotide selected from: SEQ ID NO: 44 or SEQ ID NO: 48 or SEQID NO: 51 or SEQ ID NO: 54 or SEQ ID NO: 57 or SEQ ID NO: 101 or SEQ IDNO: 159 or SEQ ID NO: 97 or SEQ ID NO: 104 or SEQ ID NO: 117 or SEQ IDNO: 143 or SEQ ID NO: 135 or SEQ ID NO: 149 or SEQ ID NO: 155 or SEQ IDNO: 46 or SEQ ID NO: 49 or SEQ ID NO: 52 or SEQ ID NO: 55 or SEQ ID NO:58 or SEQ ID NO: 102 or SEQ ID NO: 161 or SEQ ID NO: 99 or SEQ ID NO:105 or SEQ ID NO: 119 or SEQ ID NO: 145 or SEQ ID NO: 137 or SEQ ID NO:151 or SEQ ID NO: 157.

In addition to the polynucleotides encoding the antibody chains, thecloning or expression vectors of the invention carry regulatorysequences that control the expression of the antibody chain genes in ahost cell. The term “regulatory sequence” is intended to includepromoters, enhancers and other expression control elements (e.g.,polyadenylation signals) that control the transcription or translationof the antibody chain genes. Such regulatory sequences are described,for example, in Goeddel (Gene Expression Technology. Methods inEnzymology 185, Academic Press, San Diego, Calif. 1990). It will beappreciated by those skilled in the art that the design of theexpression vector, including the selection of regulatory sequences, maydepend on such factors as the choice of the host cell to be transformed,the level of expression of protein desired, etc. Regulatory sequencesfor mammalian host cell expression include viral elements that directhigh levels of protein expression in mammalian cells, such as promotersand/or enhancers derived from cytomegalovirus (CMV), Simian Virus 40(SV40), adenovirus (e.g., the adenovirus major late promoter (AdMLP)),and polyoma. Alternatively, nonviral regulatory sequences may be used,such as the ubiquitin promoter or P-globin promoter. Still further,regulatory elements composed of sequences from different sources, suchas the SRa promoter system, which contains sequences from the SV40 earlypromoter and the long terminal repeat of human T cell leukemia virustype 1 (Takebe, Y. et al., 1988 Mol. Cell. Biol. 8:466-472).

Furthermore, the cloning or expression vectors of the invention maycarry additional sequences, such as sequences that regulate replicationof the vector in host cells (e.g., origins of replication) andselectable marker genes. The selectable marker gene facilitatesselection of host cells into which the vector has been introduced (see,e.g., U.S. Pat. Nos. 4,399,216, 4,634,665 and 5,179,017, all by Axel etal.). For example, typically the selectable marker gene confersresistance to drugs, such as G418, hygromycin or methotrexate, on a hostcell into which the vector has been introduced. Selectable marker genesinclude the dihydrofolate reductase (DHFR) gene (for use in dhfr-hostcells with methotrexate selection/amplification) and the neo gene (forG418 selection).

For expression of the light and heavy chains, the expression vector(s)encoding the heavy and light chains is transfected into a host cell bystandard techniques. The various forms of the term “transfection” areintended to encompass a wide variety of techniques commonly used for theintroduction of exogenous DNA into a prokaryotic or eukaryotic hostcell, e.g., electroporation, calcium-phosphate precipitation,DEAE-dextran transfection and the like. It is theoretically possible toexpress the antibodies of the invention in either prokaryotic oreukaryotic host cells. Expression of antibodies in eukaryotic cells, forexample mammalian host cells, yeast or filamentous fungi, is discussedbecause such eukaryotic cells, and in particular mammalian cells, aremore likely than prokaryotic cells to assemble and secrete a properlyfolded and immunologically active antibody. Prokaryotic expression ofantibody genes has been reported to be ineffective for production ofhigh yields of active antibody (Boss, M. A. and Wood, C. R., 1985Immunology Today 6:12-13).

Suitable host cells for cloning or expressing vectors encodingantibodies of the invention are prokaryotic, yeast or higher eukaryoticcells. Suitable prokaryotic cells include eubacteria e.g.enterobacteriaceae such as Escherichia e.g. E. coli (for example ATCC31, 446; 31, 537; 27,325), Enterobacter, Erwinia, Klebsiella proteus,Salmonella e.g. Salmonella typhimurium, Serratia e.g. Serratiamarcescens and Shigella as well as Bacilli such as B. subtilis and B.licheniformis (see DD 266 710), Pseudomonas such as P. aeruginosa andStreptomyces. Of the yeast or fungi host cells, Saccharomycescerevisiae, Schizosaccharomyces pombe, Kluyveromyces (e.g. ATCC 16,045;12,424; 24178; 56,500), Yarrowia (EP402, 226), Pichia pastoris(EP183070, see also Peng et al (2004) J Biotechnol; 108:185-192),Candida, Trichoderma reesei (EP244234), Penicillium, Tolypocladium andAspergillus hosts such as A. nidulans and A. niger are alsocontemplated.

Although prokaryotic and yeast host cells are specifically contemplatedby the invention, preferably however, host cells of the presentinvention are higher eukaryotic cells.

Accordingly, in one aspect the present invention provides for a hostcell comprising one or more cloning or expression vectors comprising thepolynucleotides as described herein.

In another aspect, the invention provide a stably transformed ortransfected host cell comprising one or more polynucleotides asdescribed herein.

Embodiment 95: A stably transformed host cell comprising a vectorencoding a heavy chain and/or light chain of the isolated antibody orfragment thereof according to any one of embodiments 1 to 88. Preferablysuch host cells comprise a first vector encoding the light chain and asecond vector encoding said heavy chain.

Suitable higher eukaryotic host cells include mammalian cells such asCOS-1 (ATCC No.CRL 1650) COS-7 (ATCC CRL 1651), human embryonic kidneyline 293, baby hamster kidney cells (BHK) (ATCC CRL.1632), BHK570 (ATCCNO: CRL 10314), 293 (ATCC NO.CRL 1573), Chinese hamster ovary cells CHO(e.g. CHO-K1, ATCC NO: CCL 61, DHFR-CHO cell line such as DG44 (seeUrlaub et al, (1986) Somatic Cell Mol. Genet. 12, 555-556)),particularly those CHO cell lines adapted for suspension culture, mouseSertoli cells, monkey kidney cells, African green monkey kidney cells(ATCC CRL-1587), HELA cells, canine kidney cells (ATCC CCL 34), humanlung cells (ATCC CCL 75), Hep G2 and myeloma or lymphoma cells e.g. NSO(see U.S. Pat. No. 5,807,715), Sp2/0, YO.

Preferably, the mammalian host cells for expressing the binding moleculeof the invention include mammalian cell lines deficient for FUT8 geneexpression, for example as described in U.S. Pat. No. 6,946,292.

Host cells transformed with vectors encoding the binding molecules maybe cultured by any method known to those skilled in the art. Host cellsmay be cultured in spinner flasks, roller bottles or hollow fibresystems but it is preferred for large scale production that stirred tankreactors are used particularly for suspension cultures. Preferably thestirred tankers are adapted for aeration using e.g. spargers, baffles orlow shear impellers. For bubble columns and airlift reactors directaeration with air or oxygen bubbles maybe used. Where the host cells arecultured in a serum free culture media it is preferred that the media issupplemented with a cell protective agent such as pluronic F-68 to helpprevent cell damage as a result of the aeration process. Depending onthe host cell characteristics, either microcarriers maybe used as growthsubstrates for anchorage dependent cell lines or the cells maybe adaptedto suspension culture (which is typical). The culturing of host cells,particularly invertebrate host cells may utilise a variety ofoperational modes such as fed-batch, repeated batch processing (seeDrapeau et al (1994) cytotechnology 15: 103-109), extended batch processor perfusion culture. Although recombinantly transformed mammalian hostcells may be cultured in serum-containing media such as fetal calf serum(FCS), it is preferred that such host cells are cultured in syntheticserum-free media such as disclosed in Keen et al (1995) Cytotechnology17:153-163, or commercially available media such as ProCHO-CDM orUltraCHO™ (Cambrex NJ, USA), supplemented where necessary with an energysource such as glucose and synthetic growth factors such as recombinantinsulin. The serum-free culturing of host cells may require that thosecells are adapted to grow in serum free conditions. One adaptationapproach is to culture such host cells in serum containing media andrepeatedly exchange 80% of the culture medium for the serum-free mediaso that the host cells learn to adapt in serum free conditions (see e.g.Scharfenberg K et al (1995) in Animal Cell technology: Developmentstowards the 21st century (Beuvery E. G. et al eds), pp 619-623, KluwerAcademic publishers).

Binding molecules of the invention secreted into the media may berecovered and purified using a variety of techniques to provide a degreeof purification suitable for the intended use. For example the use ofisolated antibodies of the invention for the treatment of human patientstypically mandates at least 95% purity, more typically 98% or 99% orgreater purity (compared to the crude culture medium). In the firstinstance, cell debris from the culture media is typically removed usingcentrifugation followed by a clarification step of the supernatant usinge.g. microfiltration, ultrafiltration and/or depth filtration. A varietyof other techniques such as dialysis and gel electrophoresis andchromatographic techniques such as hydroxyapatite (HA), affinitychromatography (optionally involving an affinity tagging system such aspolyhistidine) and/or hydrophobic interaction chromatography (HIC, seeU.S. Pat. No. 5,429,746) are available. In one embodiment, theantibodies of the invention, following various clarification steps, arecaptured using Protein A or G affinity chromatography followed byfurther chromatography steps such as ion exchange and/or HAchromatography, anion or cation exchange, size exclusion chromatographyand ammonium sulfate precipitation. Typically, various virus removalsteps are also employed (e.g. nanofiltration using e.g. a DV-20 filter).Following these various steps, a purified (preferably monoclonal)preparation comprising at least 75 mg/ml or greater e.g. 100 mg/ml orgreater of the isolated antibody of the invention or antigen bindingfragment thereof is provided and therefore forms an embodiment of theinvention. Suitably such preparations are substantially free ofaggregated forms of antibodies of the invention.

Bacterial systems may be used for the expression of non-immunoglobulinbinding molecules described above. Bacterial systems are alsoparticularly suited for the expression of isolated antibody fragments.Such fragments are localised intracellularly or within the periplasma.Insoluble periplasmic proteins can be extracted and refolded to formactive proteins according to methods known to those skilled in the art,see Sanchez et al (1999) J. Biotechnol. 72, 13-20 and Cu pit P M et al(1999) Lett Appl Microbiol, 29, 273-277.

Accordingly, in one aspect the present invention provide a method forproducing a binding molecule which binds IL-18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex wherein the methodcomprises culturing a host cell under conditions suitable for producingthe binding molecule, wherein the host cell comprises a vector asdescribed herein.

Embodiment 95: The method of producing a human antibody or a fragmentthereof according to any one of embodiments 1 to 88, wherein the methodcomprises culturing a host cell under conditions suitable for producingthe binding molecule, wherein the host cell comprises a vector asdescribed herein.

6. Pharmaceutical Compositions

The invention provides for pharmaceutical compositions comprising thebinding molecule which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex and wherein the binding molecule isnot IL-18BP and a pharmaceutically acceptable carrier.

Embodiment 96: The pharmaceutical compositions comprising a humanantibody or a fragment thereof according to any one of embodiments 1 to88 and a pharmaceutically acceptable carrier.

The compositions can additionally contain other therapeutic agents thatare suitable for treating or preventing a human disease or disordernoted below. Pharmaceutically carriers enhance or stabilize thecomposition, or facilitate the preparation of the composition.Pharmaceutically acceptable carriers include solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents, and the like that are physiologically compatible.

A pharmaceutical composition of the present invention can beadministered by a variety of methods known in the art. The route and/ormode of administration vary depending upon the desired results. It ispreferred that administration be intravenous, intramuscular,intraperitoneal, or subcutaneous, or administered proximal to the siteof the target. The pharmaceutically acceptable carrier should besuitable for intravenous, intramuscular, subcutaneous, parenteral,spinal or epidermal administration (e.g., by injection or infusion).Depending on the route of administration, the active compound(particularly low molecular weight chemical entities) may be coated in amaterial to protect the compound from the action of acids and othernatural conditions that may inactivate the compound.

The composition should be sterile and fluid. Proper fluidity can bemaintained, for example, by use of coating such as lecithin, bymaintenance of required particle size in the case of dispersion and byuse of surfactants. In many cases, it is preferable to include isotonicagents, for example, sugars, polyalcohols such as mannitol or sorbitol,and sodium chloride in the composition. Long-term absorption of theinjectable compositions can be brought about by including in thecomposition an agent which delays absorption, for example, aluminummonostearate or gelatin.

Pharmaceutical compositions of the invention can be prepared inaccordance with methods well known and routinely practiced in the art.See, e.g., Remington: The Science and Practice of Pharmacy, MackPublishing Co., 20th ed., 2000; and Sustained and Controlled ReleaseDrug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., NewYork, 1978. Pharmaceutical compositions are preferably manufacturedunder GMP conditions. Typically, a therapeutically effective dose orefficacious dose of an antibody of the invention described herein isemployed in the pharmaceutical compositions of the invention. They aretypically formulated into pharmaceutically acceptable dosage forms byconventional methods known to those of skill in the art. Dosage regimensare adjusted to provide the optimum desired response (e.g., atherapeutic response). For example, a single bolus may be administered,several divided doses may be administered over time or the dose may beproportionally reduced or increased as indicated by the exigencies ofthe therapeutic situation. It is especially advantageous to formulateparenteral compositions in dosage unit form for ease of administrationand uniformity of dosage. Dosage unit form as used herein refers tophysically discrete units suited as unitary dosages for the subjects tobe treated; each unit contains a predetermined quantity of activecompound calculated to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of the present invention can be varied so as to obtain anamount of the active ingredient which is effective to achieve thedesired therapeutic response for a particular patient, composition, andmode of administration, without being toxic to the patient. The selecteddosage level depends upon a variety of pharmacokinetic factors includingthe activity of the particular compositions of the present inventionemployed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compositions employed, the age, sex, weight, condition,general health and prior medical history of the patient being treated,and like factors.

A physician can start doses of the antibodies of the invention employedin the pharmaceutical composition at levels lower than that required toachieve the desired therapeutic effect and gradually increase the dosageuntil the desired effect is achieved. In general, effective doses of thecompositions of the present invention, for the treatment of a fibroticdisease or disorder described herein vary depending upon many differentfactors, including means of administration, target site, physiologicalstate of the patient, whether the patient is human or an animal, othermedications administered, and whether treatment is prophylactic ortherapeutic. Treatment dosages need to be titrated to optimize safetyand efficacy. For administration with an antibody, the dosage rangesfrom about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg, of thehost body weight. For example dosages can be 1 mg/kg body weight or 10mg/kg body weight or within the range of 1-10 mg/kg. An exemplarytreatment regime entails administration once per every two weeks or oncea month or once every 3 to 6 months.

Binding molecules of the invention, especially antibodies and fragmentsthereof, are usually administered on multiple occasions. Intervalsbetween single dosages can be weekly, monthly or yearly. Intervals canalso be irregular as indicated by measuring blood levels of therapeuticprotein in the patient. In some methods, dosage is adjusted to achieve aplasma antibody concentration of 1-1000 μg/ml and in some methods 25-300μg/ml. Alternatively, antibodies of the invention can be administered asa sustained release formulation, in which case less frequentadministration is required. Dosage and frequency vary depending on thehalf-life of the antibody in the patient. In general, humanizedantibodies show longer half life than that of chimeric antibodies andnonhuman antibodies. The dosage and frequency of administration can varydepending on whether the treatment is prophylactic or therapeutic. Inprophylactic (preventative) applications, a relatively low dosage isadministered at relatively infrequent intervals over a long period oftime. Some patients continue to receive treatment for the rest of theirlives. In therapeutic applications, a relatively high dosage atrelatively short intervals is sometimes required until progression ofthe disease is reduced or terminated, and preferably until the patientshows partial or complete amelioration of symptoms of disease.Thereafter, the patient can be administered a prophylactic regime.

The pharmaceutical composition may comprise (e.g. as its soletherapeutically active ingredient) a binding molecule that binds withIL18 (e.g. specifically binds) and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex, wherein the binding molecule is not IL-18BP,wherein the binding molecule is selected from: an isolated antibody, afragment of an isolated antibody, a single variable domain antibody, abi- or multi-specific antibody, a multivalent antibody, a dual variabledomain antibody, an immuno-conjugate, a fibronectin molecule, anadnectin, a DARPin, an avimer, an affibody, an anticalin, an affilin, aprotein epitope mimetic or combinations thereof and wherein thecomposition may further comprise a pharmaceutically acceptable carrier.

Typically the composition will be in an intravenously, inhalable orsubcutaneously administrable form. In other embodiments, the compositionmaybe in lyophilized form.

Preferably, the binding molecule is an antibody, preferably a monoclonalintact antibody (e.g. human, humanised or chimeric) or a fragmentthereof as described herein.

Embodiment 97: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody or a fragment thereofthat binds IL18 (e.g. specifically binds) and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex wherein said antibodycomprises;

(a) CDRH1 of SEQ ID NO:3; (b) CDRH2 of SEQ ID NO:9; (c) CDRH3 of SEQ IDNO:5; (d) CDRL1 of SEQ ID NO:6; (e) CDRL2 of SEQ ID NO:7 and (f) CDRL3of SEQ ID NO:8.

Embodiment 98: The pharmaceutical composition according to embodiment 97wherein the antibody competes with murine antibody 125-2H for bindingIL-18.

Embodiment 99: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody or a fragment thereofthat binds IL18 (e.g. specifically binds) and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex wherein said antibodycomprises;

(a) CDRH1 of SEQ ID NO:3; (b) CDRH2 of SEQ ID NO:10; (c) CDRH3 of SEQ IDNO:5; (d) CDRL1 of SEQ ID NO:6; (e) CDRL2 of SEQ ID NO:7 and (f) CDRL3of SEQ ID NO:8.

Embodiment 100: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody or a fragment thereofthat binds IL18 (e.g. specifically binds) and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex wherein said antibodycomprises;

(a) CDRH1 of SEQ ID NO:3; (b) CDRH2 of SEQ ID NO:13; (c) CDRH3 of SEQ IDNO:5; (d) CDRL1 of SEQ ID NO:6; (e) CDRL2 of SEQ ID NO:7 and (f) CDRL3of SEQ ID NO:8.

Embodiment 101: The pharmaceutical composition according to embodiment100 wherein the antibody competes with murine antibody 125-2H forbinding IL-18.

Embodiment 102: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody or a fragment thereofthat binds IL18 (e.g. specifically binds) and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex wherein said antibodycomprises;

(a) CDRH1 of SEQ ID NO:106; (b) CDRH2 of SEQ ID NO:107; (c) CDRH3 of SEQID NO:108; (d) CDRL1 of SEQ ID NO:109; (e) CDRL2 of SEQ ID NO:110 and(f) CDRL3 of SEQ ID NO:111.

Embodiment 103: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody that binds IL18 (e.g.specifically binds) and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex wherein said antibody comprises:

-   a. a heavy chain variable domain of SEQ ID NO: 14 and a light chain    variable domain of SEQ ID NO: 16 or-   b. a heavy chain variable domain of SEQ ID NO: 25 and a light chain    variable domain of SEQ ID NO: 16 or-   c. a heavy chain variable domain of SEQ ID NO: 28 and a light chain    variable domain of SEQ ID NO: 16 or-   d. a heavy chain variable domain of SEQ ID NO: 18 and a light chain    variable domain of SEQ ID NO: 20 or-   e. a heavy chain variable domain of SEQ ID NO: 37 and a light chain    variable domain of SEQ ID NO: 20 or-   f. a heavy chain variable domain of SEQ ID NO: 40 and a light chain    variable domain of SEQ ID NO: 20 or-   g. a heavy chain variable domain of SEQ ID NO: 112 and a light chain    variable domain of SEQ ID NO: 114.

Embodiment 104: The pharmaceutical composition according to embodiment103, wherein said antibody comprises a heavy chain variable domain ofSEQ ID NO: 14 and a light chain variable domain of SEQ ID NO: 16 or aheavy chain variable domain of SEQ ID NO: 18 and a light chain variabledomain of SEQ ID NO: 20.

Embodiment 105: The pharmaceutical composition according to embodiment104 wherein the antibody competes with murine antibody 125-2H forbinding IL-18.

Embodiment 106: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody that binds IL18 (e.g.specifically binds) and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex wherein said antibody comprises:

-   -   a. a heavy chain variable domain capable of being encoded by an        isolated polynucleotide having at least 90% identity (e.g. 95%        or greater such as 96%, 97%, 98% or 99%) to an isolated        polynucleotide encoding SEQ ID NO: 14 or 18 or 25 or 28 or 37 or        40 or 112 and    -   b. a light chain variable domain capable of being encoded by an        isolated polynucleotide having at least 90% identity (e.g. 95%        or greater such as 96%, 97%, 98% or 99%) to an isolated        polynucleotide encoding SEQ ID NO: 16 or 20 or 114.

Embodiment 107: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody that binds IL18 (e.g.specifically binds) and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex wherein said antibody comprises:

-   -   a. a heavy chain of SEQ ID NO: 43 and a light chain variable        domain of SEQ ID NO: 45 or    -   b. a heavy chain of SEQ ID NO: 47 and a light chain variable        domain of SEQ ID NO: 45 or    -   c. a heavy chain of SEQ ID NO: 50 and a light chain variable        domain of SEQ ID NO: 45 or    -   d. a heavy chain variable domain of SEQ ID NO: 53 and a light        chain variable domain of SEQ ID NO: 160 or    -   e. a heavy chain variable domain of SEQ ID NO: 100 and a light        chain variable domain of SEQ ID NO: 160 or    -   f. a heavy chain variable domain of SEQ ID NO: 158 and a light        chain variable domain of SEQ ID NO: 160 or    -   g. a heavy chain variable domain of SEQ ID NO: 116 and a light        chain variable domain of SEQ ID NO: 118.

Embodiment 108: The pharmaceutical composition according to embodiment107, wherein the antibody comprises a heavy chain of SEQ ID NO: 43 and alight chain variable domain of SEQ ID NO: 45 or a heavy chain variabledomain of SEQ ID NO: 158 and a light chain variable domain of SEQ ID NO:160.

Embodiment 109: The pharmaceutical composition according to embodiment108, wherein the antibody competes with murine antibody 125-2H forbinding IL-18.

Embodiment 110: A pharmaceutical composition comprises (e.g. as its soletherapeutically active ingredient) an antibody that binds IL18 (e.g.specifically binds) and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex wherein said antibody comprises:

-   -   a. a heavy chain capable of being encoded by an isolated        polynucleotide having at least 90% identity (e.g. 95% or greater        such as 96%, 97%, 98% or 99%) to an isolated polynucleotide        encoding SEQ ID NO: 43 or 47 or 50 or 53 or 100 or 116 or 158        and    -   b. a light chain capable of being encoded by an isolated        polynucleotide having at least 90% identity (e.g. 95% or greater        such as 96%, 97%, 98% or 99%) to an isolated polynucleotide        encoding SEQ ID NO: 45 or 160 or 118.

7. Clinical Uses

It has been demonstrated that IL-18 expression is up-regulated inseveral autoimmune, cardiovascular and inflammatory diseases.

Accordingly, the binding molecules of the invention, which bind IL-18and do not bind the IL-18/IL-18 binding protein (IL-18 BP) complex andwherein the binding molecules are not IL-18BP, may be used in therapy.

Embodiment 110(a): The antibody according to any one of embodiments 1 to88 for use in therapy.

In one aspect of the present invention there is provided a method oftreating and/or preventing autoimmune diseases, including RheumatoidArthritis (RA), Systemic Onset Juvenile Arthritis (SOJA), SystemicJuvenile Idiopathic Arthritis (SJIA), Ankylosing Spondylitis, AutoimmuneInner Ear Disease (AIED), Autoimmune Lymphoproliferative Syndrome(ALPS), Behcet's Disease, Berger's Disease (IgA Nephropathy), BullousPemphigoid, Churg Strauss Syndrome, Colitis, Crohn's Disease (CD),Diabetes Type 1, Diabetes Type 2, Sjögren's Syndrome (SS),Graft-Versus-Host Disease (GVHD), Glomerulonephritis, Lupus, MultipleSclerosis (MS), Psoriasis, Rheumatic Fever, Sarcoidosis, Scleroderma,Adult Onset Still's Disease (AOSD), Systemic Lupus Erythematosus (SLE),Ulcerative Colitis, Hemophagocytic lymphohistiocytosis (HLH, also knownas hemophagocytic syndrome, macrophage activation syndrome), Familialhemophagocytic lymphohistiocytosis (FHL) and other immunodeficiencysyndromes, Giant Cell Arteritis (GCA), coronary artery disease (CAD),Coronary Vasculopathy (CV), Acute Coronary Syndromes (ACS), CongestiveHeart Failure (CHF), atherosclerosis, arteriosclerosis, myocardialinfarction (MI), Cardiorenal Syndrome (CRS), Acute Kidney injury (AKI),Diabetic Nephropathy, insulin resistance, obesity and the metabolicsyndrome (MetS), lung diseases including pulmonary sarcoidosis, inparticular pulmonary sarcoidosis pulmonary fibrosis, asthma, especiallysevere asthma, Uveitis, Geographic Atrophy Chronic Obstructive PulmonaryDisease (COPD), cystic fibrosis, Adult Respiratory Distress Syndrome(ARDS), Acute Lung Injury (ALI), Ventilator Induced Lung Injury (VILI),Pulmonary Arterial Hypertension (PAH), Alzheimer's Disease (AD) andsepsis and any combinations thereof which method comprisesadministrating to a mammalian patient a therapeutically effective amountof a binding molecule (e.g. an antibody) or pharmaceutical compositionas described herein which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex.

Embodiment 111: A method of treating and/or preventing sarcoidosis, inparticular pulmonary sarcoidosis, hemophagocytic lymphohistiocytosis(HLH), familial hemophagocytic lymphohistiocytosis (FHL), chronicobstructive pulmonary disease (COPD), Adult Onset Still's Disease(AOSD), atherosclerosis, systemic juvenile idiopathic arthritis (SJIA),severe asthma, Uveitis, Geographic Atrophy, Giant Cell Arthritis (GCA),diabetes type 1, diabetes type 2 and any combination thereof in amammalian patient which method comprises administrating to the mammalianpatient a therapeutically effective amount of a binding molecule (e.g.an antibody) or pharmaceutical composition as described herein whichbinds IL-18 and does not bind the IL-18/IL-18 binding protein (IL-18 BP)complex and wherein the binding molecule is not IL-18BP.

Embodiment 112: The binding molecules or the pharmaceutical compositionsof the invention which bind IL-18 and do not bind the IL-18/IL-18binding protein (IL-18 BP) complex and wherein the binding molecule isnot IL-18BP, for use in treating and/or preventing sarcoidosis, inparticular pulmonary sarcoidosis, hemophagocytic lymphohistiocytosis(HLH), familial hemophagocytic lymphohistiocytosis (FHL), chronicobstructive pulmonary disease (COPD), Adult Onset Still's Disease(AOSD), atherosclerosis, systemic juvenile idiopathic arthritis (SJIA),severe asthma, Uveitis, Geographic Atrophy, Giant Cell Arthritis (GCA),diabetes type 1, diabetes type 2 and any combination thereof in amammalian patient.

Embodiment 113: A method of treating and/or preventing autoimmunediseases, including Rheumatoid Arthritis (RA), Systemic Onset JuvenileArthritis (SOJA), Systemic Juvenile Idiopathic Arthritis (SJIA),Ankylosing Spondylitis, Autoimmune Inner Ear Disease (AIED), AutoimmuneLymphoproliferative Syndrome (ALPS), Behcet's Disease, Berger's Disease(IgA Nephropathy), Bullous Pemphigoid, Churg Strauss Syndrome, Colitis,Crohn's Disease (CD), Diabetes Type 1, Diabetes Type 2, Sjögren'sSyndrome (SS), Graft-Versus-Host Disease (GVHD), Glomerulonephritis,Lupus, Multiple Sclerosis (MS), Psoriasis, Rheumatic Fever, Sarcoidosis,Scleroderma, Adult Onset Still's Disease (AOSD), Systemic LupusErythematosus (SLE), Ulcerative Colitis, Hemophagocyticlymphohistiocytosis (HLH, also known as hemophagocytic syndrome,macrophage activation syndrome), Familial hemophagocyticlymphohistiocytosis (FHL) and other immunodeficiency syndromes, GiantCell Arteritis (GCA), coronary artery disease (CAD), CoronaryVasculopathy (CV), Acute Coronary Syndromes (ACS), Congestive HeartFailure (CHF), atherosclerosis, arteriosclerosis, myocardial infarction(MI), Cardiorenal Syndrome (CRS), Acute Kidney injury (AKI), DiabeticNephropathy, insulin resistance, obesity and the metabolic syndrome(MetS), lung diseases including pulmonary sarcoidosis, pulmonaryfibrosis, asthma, especially severe asthma, Uveitis, Geographic Atrophy,Chronic Obstructive Pulmonary Disease (COPD), cystic fibrosis, AdultRespiratory Distress Syndrome (ARDS)), Acute Lung Injury (ALI),Ventilator Induced Lung Injury (VILI), Pulmonary Arterial Hypertension(PAH), Alzheimer's Disease (AD) and sepsis and any combinations thereofwhich method comprises administrating to a mammalian patient atherapeutically effective amount of an antibody or pharmaceuticalcomposition as described in any one of embodiments 1 to 88 or 96 to 110which binds IL-18 and does not bind the IL-18/IL-18 binding protein(IL-18 BP) complex.

Embodiment 114: A method of treating and/or preventing sarcoidosis, inparticular pulmonary sarcoidosis, hemophagocytic lymphohistiocytosis(HLH), familial hemophagocytic lymphohistiocytosis (FHL), chronicobstructive pulmonary disease (COPD), Adult Onset Still's Disease(AOSD), atherosclerosis, systemic juvenile idiopathic arthritis (SJIA),severe asthma, Uveitis, Geographic Atrophy, Giant Cell Arthritis (GCA),diabetes type 1, diabetes type 2 and any combination thereof in amammalian patient which method comprises administrating to the mammalianpatient a therapeutically effective amount of an antibody orpharmaceutical composition as described in any one of embodiments 1 to88 or 96 to 110 which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex.

Embodiment 115: The antibody or pharmaceutical composition as describedin any one of embodiments 1 to 88 or 96 to 110 which binds IL-18 anddoes not bind the IL-18/IL-18 binding protein (IL-18 BP) complex andwherein the binding molecule is not IL-18BP, for use in treating and/orpreventing sarcoidosis, in particular pulmonary sarcoidosis,hemophagocytic lymphohistiocytosis (HLH), familial hemophagocyticlymphohistiocytosis (FHL), chronic obstructive pulmonary disease (COPD),Adult Onset Still's Disease (AOSD), atherosclerosis, systemic juvenileidiopathic arthritis (SJIA), severe asthma, Uveitis, Geographic Atrophy,Giant Cell Arthritis (GCA), diabetes type 1, diabetes type 2 and anycombination thereof in a mammalian patient.

8. Diagnostic Uses and Kits

One of the advantages of the binding molecule or isolated antibody orfragment thereof as described herein is that they bind IL-18 but do notbind the IL-18/IL-18 binding protein (IL-18 BP) complex and that theyare not IL-18BP. Hence, the binding molecule or isolated antibody orfragment thereof as described herein are capable of only detecting freeIL-18, not bound to IL-18BP. As free IL-18 is the biologically activemolecule, it becomes immediately apparent that a binding moleculecapable of recognizing only such entity may allow differentiation offree IL-18 from IL-18 bound to IL-18BP, which is inactive. Moreover, asthe binding molecules of the invention do not encompass the IL-18BP,either isolated or naturally occurring, they lend themselves not only astherapeutic agents but also as tools in many other applications such asin diagnosis or in a diagnostic kit.

Hence, in another aspect, the present invention provides for bindingmolecules or isolated antibodies or fragments thereof as describedherein which bind IL-18 and do not bind the IL-18/IL-18 binding protein(IL-18 BP) complex wherein the binding molecule is not IL-18BP for usein diagnosis or for use in a diagnostic kit.

The binding molecules or the isolated antibody or a fragment thereof asdescribed herein can be used in combination with detecting antibodies inELISA assays, western blots etc. and can be used only to detect thepresence of free IL-18 but also to measure the amount of free IL-18.

Therefore, in another aspect, the present invention provides for bindingmolecules or isolated antibodies or fragments thereof which bind IL-18and do not bind the IL-18/IL-18 binding protein (IL-18 BP) complexwherein the binding molecule is not IL-18BP for use in diagnosis or foruse in detecting and/or measuring the presence and/or the amount of freeIL-18 (i.e. IL-18 not bound to IL-18BP) in a sample.

In yet a further aspect of the present invention, there is provided amethod for detecting and/or measuring the presence and/or amount of freeIL-18 (i.e. IL-18 not bound to IL-18BP) in a sample, wherein the sampleis optionally a human sample, wherein the method comprises contactingthe sample with a binding molecule or an isolated antibody or a fragmentthereof which binds IL-18 and does not bind the IL-18/IL-18 bindingprotein (IL-18 BP) complex wherein the binding molecule is not IL-18BP.

In yet a further aspect of the present invention, there is provided amethod which comprises the steps of:

-   -   a. contacting a sample obtained from a subject (e.g. a subject        afflicted with an inflammatory disease or disorder) with the        binding molecule or the isolated antibody or a fragment thereof        as described herein which binds IL-18 and does not bind the        IL-18/IL-18 binding protein (IL-18 BP) complex wherein the        binding molecule is not IL-18BP;    -   b. measuring the level of free IL-18;    -   c. optionally, measuring the level of total IL-18 and IL-18 BP;    -   d. wherein the steps b) and c) may be carried out simultaneously        or consecutively or in inverted order (e.g. step b) before        step c) or step c) before step b).

The sample may be a sample isolated from a mammalian body, preferablyfrom a human body. The isolated sample may be:

-   -   i. from an accessible body site, for example a mucous membrane        of the nose, skin, conjunctiva, mouth or throat, anus, vagina,        urethra, cervix; and or    -   ii. comprises a fluid or semi-solid (for example a bodily fluid        or semi-solid e.g. discharge, vomit, secretion, excreta, gastric        and/or intestinal juices, sputum, blood, blood serum, plasma,        urine, tears, synovial fluid, semen, prostate fluid, saliva,        tissue homogenate or mucus); and or    -   iii. comprises a solid (e.g. stool, tissue, or biopsy sample);        and/or    -   iv. comprises a culture (e.g. macrophage culture); The sample        may be human blood or a part of human blood such as plasma.

The isolated sample may be preferably selected from human whole blood,human blood monocyte-derived macrophages and human lung macrophages.

In another aspect, the present invention further provides for adiagnostic kit comprising the binding molecule or isolated antibody or afragment thereof (which binds IL-18 and does not bind the IL-18/IL-18binding protein (IL-18 BP) complex wherein the binding molecule is notIL-18BP) and/or the complex comprising IL-18 and that binding moleculewherein the kit optionally comprises a first control compound.

A control compound is a compound that will indicate the diagnostic kitis working. A control compound may be positive or negative and it willverify that any result is valid.

The first control compound may be free IL-18 and the kit optionally maycomprise a second control compound which is murine antibody 125-2H.

In another aspect of the present invention, there is provided a medicalor diagnostic device comprising the binding molecule or the isolatedantibody or a fragment thereof (which binds IL-18 and does not bind theIL-18/IL-18 binding protein (IL-18 BP) complex) and/or the complexwherein the binding molecule is not IL-18BP comprising IL-18 and thatbinding molecule.

9. Exemplification

Sequences of the antibodies of the present invention exemplified herein,together with a sequence correlation table are described towards the endof this specification.

9.1) Materials

Recombinant human IL-18 (generated in E. coli using the method describedby Liu et al (2000) Cytokine 12(10):1519-25).

Recombinant cynomogus IL18 (generated in E. coli using the methoddescribed in U.S. Pat. No. 6,432,678) Recombinant human TNFα (R&D#210-TA)

Recombinant human IL-18 Binding Protein-IgG, Fc (hIL-18BPa-Fc, R&DSystems #119-BP-100)

Mouse anti-human IL-18 IgG, (125-2H; R&D Systems #D044-3)

Human anti-lysozyme antibody (MOR03207 Morphosys)

KG-1 cell line (ATCC #CCL-246)

Cell media: RPMI 1640 (Invitrogen #31870) supplemented with 10% FoetalBovine Serum (Invitrogen #10108-157), 1% L-Glutamine (Invitrogen#25030-03), 1% penicillin/streptomycin (Invitrogen #15140-148).

Flat bottomed, tissue-culture treated 96-well plates (Costar #3596)

Human IFN-γ DuoSet ELISA assay (R&D #DY285)

Maxisorb microtitre plates (Sigma #M9410)

Heparin (Sigma #H3393)

Recombinant human IL-12 (R&D Systems #219-IL-CF)

LPS (Sigma #L4516)

Falcon tubes (Corning #430829)

Ficoll-Paque™ Plus (GE Healthcare Life Sciences #17-1440-02)

KG-1 cells were maintained in RPMI 1640 supplemented with 10% FBS, 1%L-Glutamine and 1% penicillin/streptomycin at a density of 2×10⁵ to1×10⁶ viable cells/ml.

9.2) Antibodies and Fragments Thereof Selected for Functional Analysis

The antibodies and fragments thereof were generated by phage display.The phagemid library used is based on the HuCAL® concept (Knappik, A. atal. (2000) J Mol Biol 296, 57-86) and employs the CysDisplay™ technologyfor displaying the Fab on the phage surface (WO 01/05950). To increaseaffinity and biological activity of selected antibody fragments, L-CDR3and H-CDR2 regions were optimized in parallel by cassette mutagenesisusing trinucleotide directed mutagenesis (Virnekas, B et al. (1994)Nucleic Acids Res 22, 5600-5607), while the framework regions were keptconstant. Antibodies from different parental frameworks were selectedfor IgG conversion and functional characterization. These antibodies andfragment thereof are shown in Table 1A below.

TABLE 1A IgGs and Fabs selected for functional analysis FrameworkMOR08776 VL1/VH4 MOR10497 VL1/VH4 MOR10501 VL1/VH4 MOR10502 VL1/VH4MOR8775 VL1/VH1 MOR9441 VL1/VH1 MOR9464 VL1/VH1 MOR9465 VL1/VH1 MOR9466VL1/VH1 MOR10579 VL1/VH1 MOR10222 VL1/VH1 MOR13363 VK1/VH3 MOR13361VK1/VH3 MOR13341 VK1/VH1 MOR13342 VK1/VH1 MOR13347 VK1/VH1

Antibody MOR9464 when formulated in a liquid formulation appeared tolose potency over time. It was hypothesized that the molecule mayundergo some modifications which affected its potency. MOR9464 wasincubated at a temperature higher than physiological temperature toaccelerate the appearing of the loss of potency. By combining complexbiophysical techniques, including mass spectrometry and reverse phaseHPLC, two distinct, yet unknown, forms of MOR9464 were isolated.

A number of modifications may occur to protein when they are in theextracellular media, during in vitro manipulation and in vivocirculation. Whatever the modification is, it can widely impact theproperties relevant to protein-based pharmaceutical. Hence, it becomesparamount to understand the structure-functional relationship of thesemodifications. Exocellular modifications are modifications which occursoon after therapeutic proteins complete the trafficking pathway, reachcell surface and they are released in the extracellular mediumenvironment and incubated there during the production period. They alsoencompass modification occurring during in vitro manipulation, such aspurification and formulation in suitable buffers, or even in vivocirculation (Zhong X. and Wright J. F., Int J of Cell Biol., 2013,1-19). Proteolytic processing is the most common and well-knownexocellular modifications as its products are easily identified.Oxidation of single amino acid residues may be more difficult tocharacterize and may involve methionine residues as well as tryptophan,cysteine, histidine and tyrosine residues. N-terminal pyroglutamateformation is also a well-known exocellular modification whereby theN-terminal Glutamine or Glutamate residues can readily cyclize with itsown terminal group to form pyrrolidone carboxylic acid. Deamidation ofasparagine and aspartate residues or aspartate isomerization are alsoknown, yet not well characterized modifications.

If the exocellular modifications occur in specific regions of theantibody, they may have a deep effect on the functional characteristicsof an antibody, such as loss of potency. If the modification occurs inthe CDR or in a very near location, it may impact of the ability of theantibody to recognize and/or specifically bind its antigen.

Analyses of the MOR9464 sequence were carried out to identify potentialsite of exocellular modifications. The mutants shown in Table 1B weregenerated.

TABLE 2B MOR9464/MOR10222 mutants MOR9464_N30T MOR9464_N30A MOR9464_N30EMOR9464_N30H MOR9464_N30K MOR9464_N30Q MOR9464_N30G MOR9464_N30VMOR9464_N30Y MOR9464_N30R MOR9464_N30I MOR9464_N30L MOR9464_N30SMOR10222_E1Q MOR10222_E1Q_N30S MOR10222_E1Q_N30D MOR10222_E1Q_N30TMOR10222_E1Q_N30S_M54Y MOR10222_E1Q_N30S_M54N MOR10222_E1Q_N30S_M54I(indicated hereafter as MOR10222_N30S_M54I) MOR10222_E1Q_S31TMOR10222_E1Q_S31N MOR10222_E1Q_S31A MOR10222_E1Q_S31T_M54YMOR10222_E1Q_S31T_M54N MOR10222_E1Q_S31T_M54I MOR10222_E1Q_S31N_M54YMOR10222_E1Q_S31N_M54N MOR10222_E1Q_S31N_M54I

Asparagine 30 just flanking the H-CDR1 of MOR9464 was considered to be apossible site of modification, together with serine 31 in the H-CDR1 andmethionine 54 in the H-CDR2. Non-enzymatic deamidation of asparagine andglutamine residues to aspartate and glutamate occurs through hydrolyticreaction and is pH, temperature and ionic strength-dependent. It alsodepends on the amino acid residues preceding the asparagine andglutamine residues, with serine and threonine increasing the deamidationrates.

Most of the MOR10222_S31X mutants, with or without methionine mutation,heavily aggregated after production. Hence these mutants were notcharacterized any further.

9.3) Determination of Affinities Using Solution Equilibrium Titration(SET)

For K_(D) determination by solution equilibrium titration (SET), monomerfractions of antibodies were used (analyzed by analytical SEC).

Affinity determination in solution was basically performed as describedin the literature (Friguet B at al. (1985) J Immunol Methods;77(2):305-19). In order to improve the sensitivity and accuracy of theSET method, it was transferred from classical ELISA toelectro-chemiluminescent (ECL)-based technology (Haenel C et al. (2005)Anal Biochem; 339(1):182-4.).

Goat-anti-human (Fab)₂ fragment specific antibodies (Dianova) werelabeled with MSD SULFO-TAG™ NHS-Ester (Meso Scale Discovery,Gaithersburg, Md., USA) according to the manufacturer's instructions.Experiments were carried out in polypropylene microtiter plates andphosphate-based assay buffer. Unlabeled human IL-18 was seriallydiluted, starting with a concentration at least 10 times higher than theexpected K_(D). Wells without antigen were used to determine B_(max)values; wells with assay buffer were used to determine background. Afteraddition of a constant amount of antibodies or fragment thereof (e.g. 10pM final concentration in 60 μl final volume), the mixture was incubatedover night at RT. The applied antibodies or fragments thereofconcentration was similar to or below the expected K_(D).

Streptavidin MSD plates were blocked with BSA-containing phosphatebuffer over night. After blocking of the plate, biotinylated human IL-18was added and incubated for 1 h at RT. Subsequently the equilibratedsamples were transferred to those plates and incubated for a short timeat RT. After washing, MSD SULFO-TAG labeled detection antibody (goatanti-human (Fab)₂) was added to the MSD plate and incubated for a shorttime at RT.

After washing the plate and adding MSD Read Buffer T with surfactant,electrochemi-luminescence signals were detected using a Sector Imager6000 (Meso Scale Discovery, Gaithersburg, Md., USA).

Data was evaluated with XLfit (IDBS) software applying customizedfitting models. For K_(D) determination of Fab molecules the followingfit model was used (according to Haenel et al., 2005), modifiedaccording to Abraham et al, 1996):

$y = {B_{\max} - \left( {\frac{B_{\max}}{{2\lbrack{Fab}\rbrack}_{t}}\left( {\lbrack{Fab}\rbrack_{t} + x + K_{d} - \sqrt{\left( {\lbrack{Fab}\rbrack_{t} + x + K_{D}} \right)^{2} - {4{x\lbrack{Fab}\rbrack}_{t}}}} \right)} \right)}$

[Fab]_(t): applied total Fab concentration

x: applied total soluble antigen concentration (binding sites)

B_(max): maximal signal of Fab without antigen

K_(D): affinity

In principle the same protocol was applied to determine K_(D) values forantibodies and fragments with the following differences: wholeantibodies were added to the dilution series of antigen, andequilibrated over night at RT. Subsequently, the samples were treated asdescribed above.

For data evaluation i.e. K_(D) determination of IgG molecules thefollowing fit model for IgG was used (modified according to Piehler etal., 1997):

$y = {\frac{2B_{\max}}{\lbrack{IgG}\rbrack}\left( {\frac{\lbrack{IgG}\rbrack}{2} - \frac{\left( {\frac{x + \lbrack{IgG}\rbrack + K_{D}}{2} - \sqrt{\frac{\left( {x + \lbrack{IgG}\rbrack + K_{D}} \right)^{2}}{4} - {x\lbrack{IgG}\rbrack}}} \right)^{2}}{2\lbrack{IgG}\rbrack}} \right)}$

[IgG]: applied total IgG concentration

x: applied total soluble antigen concentration (binding sites)

B_(max): maximal signal of IgG without antigen

K_(D): affinity

Affinities of the anti-IL-18 antibodies or fragments thereof weredetermined in solution using the assay conditions described above andare indicated in Table 2A (column 2). Where IL-18 from cynomolgus monkey(cm IL-18) was also tested, the resulting K_(D) are indicated inbrackets.

9.4) Inhibition of IFN-γ Release from IL-18 Stimulated Human PBMCs

Peripheral blood mononuclear cells (PBMCs) were freshly isolated fromheparinized human whole blood using standard techniques. Briefly,diluted blood containing Heparin (15 U/ml) as an anti-coagulant waslayered onto Ficoll-Paque™ Plus and centrifuged (800×g, 20 minutes, 18°C.). The PBMCs were collected from the plasma:Ficoll interface andwashed twice (600×g, 10 minutes, 4° C.), before re-suspending in medium.

PBMCs were re-suspended in media and applied to a 96 well cell cultureplate to give a final cell density of 1.5×10⁶/ml (in a final assayvolume of 200 μl). To ensure the antibodies and fragment thereofaccording to the invention recognised native IL-18, which may beglycosylated, PBMCs were stimulated with LPS and IL-12 which inducesIFN-γ via IL-18-dependent autocrine feedback.

Cells were stimulated with 3 nM human recombinant IL-18 plus IL-12 (1ng/ml) or LPS (3 μg/ml) plus IL-12 (10 ng/ml) to induce secretion ofnative IL-18 protein (all final concentrations). In the case ofrecombinant human IL-18, the concentration selected was pre-determinedas an approximate ECK) in this assay. In the case of native IL-18, LPSstimulates production of IL-18, whilst IL-12 increases IL-18 receptorexpression (Yoshimoto T et al. (1998) J Immunol; 161(7):3400-7). Theextent of IL-18-dependency in this assay was determined via inclusion ofthe highly specific protein, IL-18 Binding Protein-IgG, Fc(hIL-18BPa-Fc) as a positive control (dotted line in FIGS. 3 (A-D).

To assess their potency and efficacy, the antibodies and fragmentsthereof were pre-equilibrated for 30 min with either IL-12 and LPS(native conditions) or IL-12 and IL-18 (recombinant conditions) prior toapplying to cells with a final concentration between 0.1 and 300 nM.Anti-lysozyme control antibody MOR03207 was used as a negative control.

From each treatment group, a mean±SEM value (where n=3 or more) wasdetermined from n=2-5 wells.

Cells were incubated at 37° C., 5% CO₂ for 24 hours after treatment,after which time the supernatant was collected by centrifugation (1200rpm for 5 min) and stored at −20° C. for subsequent analysis. IFN-γprotein levels were assessed using ELISA as per the manufacturer'sinstructions.

All antibodies and fragments thereof dose-dependently inhibited IFN-γproduction with a similar level of efficacy achieved at the highestconcentrations as that seen for recombinant IL-18BPa-Fc (Table 2Acolumns 5-6, Table 2B column 2 and FIGS. 3 (A-D) where efficacy of theIL-18BPa is denoted by dotted line). The control antibody, MOR03207(anti-lysozyme control antibody) did not inhibit the IL-18 response.Germlining antibodies, (e.g. MOR10579 from MOR09441 and MOR10222) fromMOR09464 did not significantly alter their neutralizing capacity.Mutants of MOR9464 and MOR10222, in particular MOR9464_N30K andMOR10222_N30S_M54I did have comparable neutralizing capacity to the wildtype antibodies.

TABLE 2A K_(D) values of IL-18 affinity and IC₅₀ values (nM) forinhibition of human recombinant (hr) IL-18 (1 nM) and nativeIL-18-induced IFNγ release K_(D) (SET, pM) KG-1 KG-1 PBMC PBMC Hu IL-18Hu IL-18 Cyno IL-18 Hu IL-18 Native IL-18 MOR08776 38  18   ND 50.8  5.9MOR010497 2 0.3 0.3 5.8 ± 2.2 1.8 ± 0.6 MOR010501 1 0.3 0.2 6.3 ± 1.51.3 ± 0.5 MOR010502 4 0.4 0.2 6.3 ± 0.7 2.3 ± 1.1 MOR08775 65  68   22  9.2 0.7 MOR09465 6 0.3 1.8 0.6 0.89 ± 0.3  MOR09466 5 0.4 5.6 1.2 2.1 ±0.6 MOR09441 1 0.6 ± 0.1 16.4 ± 7.1  2.5 ± 1.0 0.45 ± 0.1  MOR09464 30.8 ± 0.2 8.3 ± 1.6 1.3 ± 0.5  0.2 ± 0.03 MOR010579 N/A 0.8 ± 0.2 9.8 ±5.3 1.9 ± 0.6  0.3 ± 0.02 MOR010222 N/A 0.7 ± 0.1 6.6 ± 2.0 2.3 ± 0.50.22 ± 0.03 IL-18BP-FC N/A 0.3 ± 0.1 0.4 ± 0.1 1.0 ± 0.5 0.17 ± 0.02MOR09464_N30K  2 ± 1 (20 ± 10) 0.36 ± 0.15 4.7  0.2 ± 0.14MOR1022_N30S_M54I 3 ± 2 (30 ± 0) 4.21 ± 0.14  8.12 0.05 ± 0.03 MOR1336310 ± 0 (10 ± 0)  0.49 ± 0.9*  0.28 ± 0.16^(#) 0.18 ± 0.1  MOR13361 7 ± 3(6 ± 1)   0.41*  0.13^(#)  0.5 ± 0.01 MOR13341 3 ± 2 (30 ± 0)  0.13 ±0.08*   2.8 ± 0.26^(#) 0.8 ± 0.7 MOR13342 N/A  0.11 ± 0.03*  2.9^(#) 0.5± 0.4 MOR13347 N/A  0.12 ± 0.07*  2.8 ± 0.3^(#) 0.5 ± 0.4 N/A = notavaiable; *human IL-18 concentration = [0.5 nM]; ^(#)cyno IL-18concentration = [0.2 nM]

TABLE 2B IC₅₀ values (nM) for inhibition of human recombinant (hr) IL-18(1 nM), native IL-18-induced IFNγ release KG-1 PBMC Identification HuIL-18 Native IL-18 MOR1022_N30S 3.3 ± 2.1 0.05 ± 0.00 MOR1022_N30S_M54Y4.8 ± 0.3 0.09 ± 0.01 MOR1022_N30S_M54N 5.2 ± 4.4 0.03 ± 0.02MOR1022_N30S_M54I 4.2 ± 1.4  0.05 ± 0.03* MOR09464_N30T 0.31 ± 0.02 0.19± 0.09 MOR09464_N30E 0.49 ± 0.05 0.13 ± 0.05 MOR09464_N30H 0.23 ± 0.110.10 ± 0.05 MOR09464_N30K 0.36 ± 0.15 0.20 ± 0.14 MOR09464_N30Q 0.22 ±0.03 0.07 ± 0.14 MOR09464_N30G 0.54 ± 0.06 0.11 ± 0.07 MOR09464_N30V0.29 ± 0.03 0.11 ± 0.02 MOR09464_N30Y 0.23 ± 0.01 0.03 ± 0.01MOR09464_N30R 0.47 ± 0.49 0.04 ± 0.01 MOR09464_N30I 0.32 ± 0.09 0.11 ±0.06 MOR09464_N30L 0.23 ± 0.03 0.05 ± 0.04 (n = 2; n = 3 Mean +/− SD)Mean +/− SEM *n = 2, Mean +/− SD

9.5) Inhibition of IFN-γ Release from IL-18 Stimulated KG-1 Cells

Anti-IL-18 antibodies and fragments thereof were analyzed for theirability to inhibit IFN-γ release from IL-18-stimulated KG-1 cells. Inthe absence of an antagonist, IL-18 promotes maximal stimulation ofIFN-γ release in the presence of a co-stimulus such as TNFα and IL-12through upregulation of IL-18 receptors (Nakamura S et al. (2000)Leukemia; 14(6):1052-9). Inhibitory activity of the antibodies andfragments was assessed against 1 nM of either human or cynomolgus IL-18(unless otherwise specified), predetermined as an EC₈₀ in thiscell-based system.

KG-1 cells were re-suspended in culture media and applied to a 96 wellcell culture plate to give a final cell density of 0.3×10⁶/ml (in afinal assay volume of 200 μl). Cells were stimulated with 1 nM human orcynomolgus recombinant IL-18 plus TNFα (final concentration 20 ng/ml).The concentration of recombinant IL-18 selected was pre-determined as anapproximate EC₈₀ in this assay.

To assess the potency and efficacy of these antibodies, the antibodiesand fragments thereof were pre-equilibrated for 30 min with TNFα andIL-18 (human or cynomolgus) prior to applying to cells with a finalconcentration between 0.1 and 300 nM. Anti-lysozyme control antibodyMOR03207 was used as a negative control.

From each treatment group, a mean±SEM value (where available) wasdetermined from n=2-5 wells.

Cells were incubated at 37° C., 5% CO₂ for 24 hours after treatment,after which time the supernatant was collected by centrifugation (885×gfor 5 min) and stored at −20° C. for subsequent analysis. IFN-γ proteinlevels were assessed using ELISA as per the manufacturer's instructions.

All antibodies and fragments thereof dose-dependently inhibited IFN-γproduction with full inhibition achieved at the highest concentrations.For affinity-matured antibodies, IC₅₀ values ranged from 0.3-0.8 nMagainst 0.8 nM human IL-18, therefore representing a 1:1 molar ratioantibody:antigen or better. VH4_3b framework antibodies (MOR08776,MOR010501, MOR010502) demonstrated similar potency against both humanand cynomolgus IL-18, whilst VH1A_3 framework antibodies were slightlyless potent against cynomolgus IL-18 (Table 2, columns 3 and 4). Thecontrol antibody, MOR03207 did not inhibit the IL-18 response.

9.6) IL-18 Induced INF-γ Release in Human Whole Blood

The whole blood assay incorporates both the presence and function of theendogenous IL-18BP since the IL-18BP is primarily generated by thespleen and is present systemically at levels of around 10-20 ng/ml inhealthy individuals. IL-18BP binds with high affinity to IL-18 andneutralizes its activity, and therefore might represent a sink forantibodies that recognize accessible epitopes of this complex.

The activity of the anti-IL-18 antibodies was assessed in heparinizedhuman whole blood taken from a healthy volunteer. All reagents wereprepared at twenty-fold the final desired concentration using serum freemedium. Cells were stimulated with either 7 nM human recombinant IL-18plus IL-12 (1 ng/mL) or LPS (10 μg/mL) plus IL-12 (10 ng/mL) to inducesecretion of native IL-18 protein (all final concentrations). In thecase of recombinant human IL-18, the concentration selected waspre-determined as an approximate ECK) in this assay. In the case ofnative IL-18, LPS stimulates production of IL-18, whilst IL-12 increasesIL-18 receptor expression and the extent of IL-18-dependency isdetermined via inclusion of recombinant human IL-18 Binding Protein-IgG,Fc (hIL-18BPa-Fc) as a positive control.

To assess the potency and efficacy of these antibodies, the anti-IL-18antibodies and fragments thereof were pre-equilibrated for 30 min withthe relevant stimulus prior to applying to cells with a finalconcentration between 0.1 and 1000 nM. Anti-lysozyme control antibodyMOR03207 was used as a negative control.

The stimulus mixture (+/−antibody or fragment) was added to each well ofa 96-well tissue culture sterile microtitre plate as 30 μL and 170 μLheparinised whole blood was then added to each well, such that the finalvolume in each well was 200 μl. The plates were then returned to ahumidified incubator (37° C.). After 24 h, the plates were centrifuged(885×g, 4° C., 5 min) and the supernatants removed and assayed forhIFN-γ production using a commercially available ELISA kit. Final datawas derived as a mean±standard error of mean from 3-5 healthy humandonors.

All antibodies and fragments dose-dependently inhibited IFNγ productionagainst either stimulus. Full inhibition of the recombinantIL-18-induced response was achieved at the highest concentrations,whilst similar efficacy was observed against LPS/IL-12 stimulation asthat for the IL-18BPa-Fc (Table 3; FIGS. 7(A-E) and 8(A-E)). GermliningMOR09441 and MOR09464 did not significantly alter their neutralizingcapacity. The control antibody, MOR03207 did not inhibit the IL-18response. Selected antibodies were also assessed for their ability toinhibit recombinant cynomolgus IL-18 bioactivity (7 nM) in whole bloodfrom the cynomolgus monkey. MOR09441, MOR09464, MOR09465, MOR09466 alldose-dependently inhibited IFN-γ production with full inhibitionobserved at the highest concentrations. Observed IC₅₀ values were 110±36nM, 51±8 nM, 55±3 nM, 179±30 nM, respectively.

TABLE 3 IC₅₀ values (nM) for inhibition of IL- 18-induced IFN-γ releasein whole blood Human whole blood stimulated Human whole with recombinantblood stimulated Identification human IL-18 (7 nM) with LPS/IL-12MOR010497 16.1 ± 6.6  107.9 ± 8.3  MOR010501 20.3 ± 5.6  101.3 ± 44 MOR010502 17.9 ± 5.0  110.4 MOR09465 4.2 ± 1.0 19.4 ± 7.8 MOR09466 5.6 ±0.5 18.5 ± 1.6 MOR09441  8 ± 2.1 16.3 ± 5.4 MOR09464 3.6 ± 0.4  9.8 ±3.2 MOR010579 5.6 ± 1.4 16.7 ± 7.0 MOR010222 4.7 ± 1.5 21.6 ± 9.9IL-18BP-Fc 5.7 ± 2.0 12.1 ± 6.2 MOR13363 29.4  91.7 ± 66.9 MOR1336115.72 ± 5.5   39.4 ± 13.5 MOR13341 5.6 ± 0.4 19.6 ± 5.3 MOR13342 6.6 ±1.0 21.5 ± 6.7 MOR13347  3.5  8.3 MOR1022_N30S 58.4 ± 14.4 16.9 ± 7.4MOR1022_N30S_M54Y 34.3 ± 16.6 18.8 ± 6.5 MOR1022_N30S_M54N 46.5 ± 9.9 16.1 ± 5.1 MOR1022_N30S_M54I 47.3 ± 15.1  9.3 ± 4.4 MOR09464_N30T 12.46± 2.67  20.76 ± 5.74 MOR09464_N30E 12.88 ± 2.79  19.16 ± 6.67MOR09464_N30H 11.93 ± 2.14   22.98 ± 11.04 MOR09464_N30K 8.62 ± 2.6217.09 ± 3.64 MOR09464_N30Q 8.10 ± 1.00 13.40 ± 2.91 MOR09464_N30G 25.14± 5.26  15.07 ± 2.80 MOR09464_N30V 12.68 ± 1.86  19.60 ± 4.90MOR09464_N30Y 12.39 ± 1.59  17.85 ± 5.12 MOR09464_N30R 10.07 ± 1.32 11.03 ± 1.26 MOR09464_N30I 11.42 ± 2.37  13.15 ± 4.53 MOR09464_N30L 9.85± 1.64 15.46 ± 2.29 n = 3-4 n = 3-4 Mean +/− SEM Mean +/− SEM * n = 2,Mean +/− SD

Mutants of MOR9464 and MOR10222, in particular MOR9464_N30K andMOR10222_N30S_M54I did have comparable neutralizing capacity to the wildtype antibodies.

9.7) ELISA Binding of Anti-IL-18 Antibodies to the IL-18-IL-18BP Complex

To confirm that the anti-IL18 antibodies or fragments thereof describedherein do not recognise IL-18 bound to the IL-18BP (IL-18/IL-18BPcomplex) IL-18 was incubated with IL-18BP (rhIL-18BP/Fc, R&D Systems,Cat #119) in a molar excess. Anti-IL18 antibodies and fragments wereadded as described below and binding to the complex was detected. Ingeneral, signals at high concentrations of the anti-IL-18 antibodies andfragments are detected where they bind accessible epitopes on theantigen different to the IL-18BP (i.e. recognize the IL-18/IL-1BPcomplex). In a first setup, control antibodies and anti-IL18 antibodiesand fragments were diluted using PBST/0.5% BSA and added to thebiotinylated IL-18/IL18-BP complex (incubation in polypropylene platesfor 30 min at RT and shaking gently). Control antibodies wereMOR03207(anti-Iysozyme) as negative control, MOR08741 as well as mouse125-2H, an anti-human IL-18 mouse IgG, as positive control antibodies(they both recognises IL-18/IL-18BP complex) (Argiriadi M A et al.(2009) J Biol Chem; 284(36):24478-89). The whole complex was capturedvia the biotin moieties onto NeutrAvidin plates, which were blocked o/nwith 1× ChemiBlocker-PBS.

Plates were washed 5× with PBST and incubated for 1 h with 20 μl/welldetection antibody anti-Fab-AP—goat anti human IgG, F(ab)2 fragmentspecific (Jackson Immuno Research, 109-055 097, lot: 69655) oranti-mouse IgG (whole molecule)-AP (SIGMA, #A4312) both diluted 1:5000in 0.5% BSA/0.05% Tween20/1×PBS. Plates were washed 5× with TBST, 20 μlAttoPhos (Roche) solution (1:5 diluted in ddH2O) were added andfluorescence was measured at the Tecan Reader.

For anti-IL-18 antibodies MOR8775 and MOR8776 which do not bind theIL-18/IL-18BP complex, binding signals were significantly decreased orno signals were observed (FIG. 9(A)). In comparison, a strong signal wasobserved in the presence of mouse 125-2H supporting the reported abilityof this control antibody to bind an epitope distinct from that of theIL-18BP. Similarly, a dose-dependent signal was observed in the presenceof control antibody MOR08741.

In a second setup, the experiment was done in a similar way, except forusing unbiotinylated hu IL-18. The IL-18/IL-18BP complex was capturedonto a Maxisorp plate via the Fc tag of the rhIL-18BP/Fc using a goatanti-hu IgG (Fc gamma fragment specific, Jackson ImmunoResearch#109-005-098). A concentration-dependent signal was observed forcontrols MOR08741, again highlighting the ability of this antibody torecognise the IL-18/IL-18BP complex. In comparison, no signal wasobserved for MOR09441, MOR09464, MOR09465, MOR09466, confirming thatthey do not bind the IL-18/IL-18BP complex.

9.8) Epitope Binning of Anti-IL18 Antibodies and Fragments Via ELISA

For epitope binning two set-ups were used. For the first set-up, anantibody fragment (Fab A) was titrated and incubated with biotinylatedhuman IL-18. Fab A was tested with a constant concentration of anantibody B (IgG B). As a positive control, Fab A was analyzed withitself in the IgG format. NeutrAvidin plates (Thermo Scientific Cat#15402) were blocked with Chemiblocker (1:1 diluted with PBS) andincubated for 2 h at RT (or over night at 4° C.). Next day plates werewashed 2× with PBST and Fabs were titrated from 500 nM down to 2 nM inPBS buffer containing 10 nM final concentration biotinylated humanIL-18. The complex of antibody fragments and biotinylated IL-18 wasadded to the NeutrAvidin plates and incubated for 1 hour. After washing3× with PBST IgG B was added at 20 nM to the corresponding wells of theNeutrAvidin plates. After 20 minutes incubation, and 3×PBST wash,detection antibodies anti-Fc-AP—goat anti human IgG, Fc gamma-chainspecific (Jackson Immuno Research, 109-055 098) and anti-mouse IgG(whole molecule)-AP (SIGMA, #A4312, Lot: 067K4863) were added diluted1:5000 in 0.5% BSA/0.05% Tween20/1×PBS.

Plates were washed 5× with PBST, 20 μl AttoPhos solution (1:5 diluted inddH2O) was added and plates were measured at the Tecan Reader.Fluorescence emission at 535 nm was recorded with excitation at 430 nm.

In general, signals could only be obtained when the IgG B was able tobind to accessible epitopes on the antigen which are different to thetested Fab A (i.e. antibody binding to a different epitope). Incontrast, for antibodies with partially overlapping or identicalepitopes, binding signals were significantly decreased in comparison tocontrols.

The second ELISA set-up was performed on Maxisorp plates. Fab A wascoated to different concentrations in PBS and incubated o/n at 4° C.Next day plates were washed 3× with PBST and blocked for 2 hours at RTwith 5% MPBST (100 μl/well). After a 3×PBST washing step, human IL-18was added at a constant concentration for 1 h. Plates were washed 3×with PBST and biotinylated antibody fragment (biotinylated Fab B) wastitrated (max. conc. 5 μg/mL, 10 μg/mL or 20 μg/mL). For complexformation of Fab A with IL-18 and biotinylated Fab B, plates wereincubated for 1 h, subsequently washed 3× with PBST and detectionantibody ZyMax Streptavidin-Alkaline Phosphatase (Zymed, Cat. No.43-8322, Lot 51102099) was diluted 1:2,000 in PBST and 20 μL/well wasadded to the ELISA plates. Plates were washed 5× with TBST, 20 μLAttoPhos (Roche) solution (1:5 diluted in ddH2O) were added and plateswere measured at the Tecan Reader.

In this case, as for the first set-up, signals could only be obtainedwhen the biotinylated Fab B was able to bind to accessible epitopes onthe antigen which are different to the tested Fab A (i.e. antibodybinding a different epitope).

As shown in FIG. 10A, antibodies MOR8775 compete with murine antibody125-2H (black cell filling) whilst MOR8776 does not compete with 125-2Hfor binding to IL-18 (no cell filling). Both antibodies do compete withIL-18BP-Fc for binding IL-18 (black cell fillings). Neither MOR8775 norMOR8776 compete with ABT325 antibody (U.S. patent application Ser. Nos.09/780,035 and 10/988,360). Depending on the experiments settings,MOR8775 and MOR8776 compete with each other (striped cell filling) In asecond set of experiments antibodies MOR9464, MOR9464_N30K,MOR10222_N30S_M54I and MOR13341 were tested for their ability to bindIL-18 in the presence of any of these antibodies, IL-18BP-Fc, ABT325 andmurine antibody 125-2H using the Proteon XPR36 instrument, a surfaceplasmon resonance (SPR) based, real-time label-free biosensor. Prior toanalysis the integrity of the proteins was confirmed-, and theconcentration assessed by LC-MS.

All antibodies and IL18BP-Fc were immobilized on the interaction spotsof a GLC sensor chip by standard amine coupling. In a first step IL18was injected as analyte 1 and in a second step either an anti-IL18antibody or IL18BPa-Fc was injected as analyte 2.

Ligands to be immobilized were prepared at a concentration of either 20μg/mL (antibodies) or 10 μg/mL (IL18BP-Fc) in 10 mM Acetate Buffer, pH4.0. Analytes were diluted in PBS buffer (TEKNOVA) containing 0.005%Tween 20, the same buffer solution was used as running buffer for theinstrument. Analyte 1 (25 nM IL18) was injected at a flow rate of 50μL/min for 180 seconds with a dissociation time of 60 seconds. Analyte 2(25 nM IL18BPa-Fc or anti-IL18 antibody) was injected at a flow rate of50 μL/min for 180 seconds with a dissociation time of 180 seconds.Regeneration was performed with 10 mM Glycine, pH 1.5 at a flow rate of100 μL/min for 20 seconds. Two independent sets of experiments wereperformed. Data evaluation was based on whether the sensogram signalincreased upon injection of analyte 2 compared to the signal ofanalyte 1. If the signal increased it was concluded that the immobilizedligand and injected analyte 2 recognized different epitopes. If thesignal did not increase it was concluded the two shared the same oroverlapping epitopes.

As shown in FIG. 10B, antibodies MOR9464, MOR9464_N30K,MOR10222_N30S_M54I and MOR13341 compete with each other (black cellfillings), with IL-18BP-Fc and with murine antibody 125-2H. None ofthese antibodies compete with ABT325 (white cell fillings).

Antibodies such as MOR9464_N30K and MOR10222_N30S_M54I which show todually compete with the IL-18BP-Fc and with murine antibody 125-2Happear to have the advantage of not only binding free IL-18, not boundto the natural inhibitor IL-18BP-Fc, but also appear to have thepotential to prevent binding of IL-18 to the IL-18Rα/β. As described inWu et al. (Wu C. et al., J. Immunol. 2003, 170: 5571-5577), IL-18R13does not appear to bind IL-18 alone, but form a functional high affinityreceptor complex with IL-18Rα that is able to signal in response toIL-18. Biochemical data combined with epitope mapping of 125-2H on IL-18have revealed that the C-terminal 17 amino acids of the human IL-18 arecritical for signal transduction through the heterodimeric receptor.Hence, antibodies capable of binding within the 125-2H epitope andequally competing with the IL-18 BP for binding IL-18 appear to have thepotential of preventing IL-18-dependent pathway activation not onlythrough blocking the binding to IL-18Rα but also through blocking thebinding of IL-18 to IL-18Rα/β complex. As it will be apparent in section9.11, antibody MOR9464_N30K binds, among others, amino acids Glu177 andLeu180 which are within the 17-amino acid stretch described in Wu et al.

9.9) IL-18 Epitope Mapping by Modelling

Human IL-18 from SwissProt entry Q14116 was used to search forstructural information in the PDB (Berman H. M et al (2000) Nucl AcidsRes; 28:235-242). Three structures were found with the code 1J0S (Kato Zet al (2003) Nat Struct Biol; 10:966), 2VXT (Argiriadi M A et al (2009)J Biol Chem; 284:24478) and 3F62 (Krumm B et al (2008) Proc Natl AcadSci USA; 105:20711). 1J0S is the NMR structure of human IL-18. 2VXT isthe crystal structure of the engineered human IL-18 in complex withmouse 125-2H antibody fragment at 1.49 Å resolution and 3F62 is thecrystal structure of the engineered human IL-18 in complex with thepoxvirus IL-18 binding protein at 2.0 Å resolution.

Three binding sites on IL-18 have been identified by mutationalanalysis. Two of them, site 1 and 2, are important for binding IL-18Rαand the third one, site 3, for binding IL-18R13 (Kato Z. et al. (2003)Nat. Struct. Biol, 10:966). Site 2 is also important for bindingIL-18BP.

A structural complex between IL-1β and IL-1R1 is also available (1ITB,Vigers G. P et al (1997) Nature, 386:190). The structure of the complexbetween IL-1β and IL-1R1 shows that IL-1β has two binding sites for thereceptor, site 1 and 2 (FIG. 11(A)). Since no structure is available forthe complex between IL-18 and IL-18Rα, the crystal structure of IL-1β incomplex with IL-1R1 was used as template for protein modelling (PDB code11 TB). The model of IL-18Rα was built by using the structure of IL-1R1as template. The crystal structure of IL-18 (pdb code 2VXT, Argiriadi M.A et al (2009) J. Biol. Chem. 284:24478) and the modelled structure ofIL-18Rα were structurally superposed to the complex IL-1β/IL-1R1 and theIL-18/IL-18Rα model so obtained was refined to obtain the finalstructural model (FIG. 11(B)). MOE v2009.1 (Chemical Computing GroupInc.) is the software used for modeling the complex between IL-18 andIL-18Rα. The Homology Model panel has been used to build the model ofIL-18Rα, by selecting AMBER99 forcefield and the default panelparameters. The energy minimization panel in MOE has been used for therefinement of the final IL-18/IL-18Rα complex.

The overall structure of human IL-18 shows similarity with that ofIL-1β. In particular, the RMSD (root mean square deviation) betweenIL-18 and IL-1 β Cα atoms in secondary structure elements indicates thatthey are related proteins belonging to the same structure class (Kato Z.et al. (2003) Nat. Struct. Biol, 10:966). A comparison between thestructures of IL-1β and IL-18 revealed that site 1 and 2 in IL-18correspond to site 1 and 2 in IL-1β. In FIG. 11(B), sites 1 and 2 areshown in the model of the complex between IL-18 and IL-18Rα, site 3 isalso shown. Site 3 is reported as the site of interaction for IL-18Rβ(Kato Z. et al. (2003) Nat. Struct. Biol, 10:966).

The IL-18 binding site for IL-18BP has been somehow identified byalanine mutations and by the X-ray crystal structure of a poxvirusIL-18BP in complex with IL-18 (Krumm et al (2008) Proc Natl Acad SciUSA; 105(52):20711-20715). This putative binding site also correspondsto the region on IL-18 that has been identified as site 2, one of thetwo regions of interaction of IL-18 with IL-18Rα.

9.10) IL-18 Epitope Mapping by Hydrogen/Deuterium Exchange MassSpectrometry (H/DxMS)

Hydrogen/Deuterium exchange Mass Spectrometry was used to probe humanIL-18 for information regarding the epitope for MOR9464. H/DxMS mappingrelies upon the mass differences between “normal” hydrogen atoms and the“heavy” isotope deuterium which also comprises a neutron in addition tothe single proton present in the normal hydrogen nucleus.

Upon transfer from water to a deuterium based solvent system (heavywater), a protein will experience an increase in mass as the amidehydrogen on the protein's backbone become gradually replaced withdeuterons (heavier isotope of hydrogen). The likelihood of ahydrogen/deuterium exchange event is largely determined by proteinstructure and solvent accessibility. The H/DxMS technology is used tomeasure relative hydrogen/deuterium exchange and as a consequenceprotein structure and solvent accessibility.

When a protein binding partner binds to an antibody (e.gantigen/antibody interaction) experimentally observable changes in itsexchange rate may be observed. Surface regions that exclude solvent uponcomplex formation exchange much more slowly. Solvent excluded regionsare useful for deducing the location of a binding site. In the case ofan antigen-antibody interaction, changes in the rate of deuteriumexchange might highlight the location of the epitope, but also any otherperturbation resulting from the binding of the antibody to the antigen.For example, a decrease in the amount of deuterium uptake in the antigenat a given exchange time after antibody binding might represent eitherincreased protection due to direct binding of the antibody to thisregion or indirect perturbation of the structure (allosteric changes)because of antibody binding. These two effects cannot be distinguishedvery easily, though the strongest effect observed is often attributed todirect protection from the antibody.

The location of decreased deuteron incorporation after antibody bindingmay be deduced by digestion of the target protein followinghydrogen/deuterium exchange (e.g. with a suitable enzyme such as pepsin)and then mass spectrometry to determine the mass of the relevantfragments.

Triplicate control experiments were carried out on 316 pmol IL-18antigen at deuterium exchange times of 3 and 25 minutes by diluting astock solution of IL-18 with 95% deuterated PBS buffer to aconcentration of 83.6% D. Deuterium exchange was quenched with quenchingbuffer (6M Urea and 1M TCEP). After quenching, the vial was analyzed byon-line pepsin digestion/LC-MS analysis. On-line pepsin digestion wasperformed using Life Science's Poroszyme immobilized pepsin packed into2.0×20 mm column, and LC separation was performed on a Thermo C18BioBasic column (1.0×50 mm) at a flow rate of 100 μL/min using a fastgradient so all peptides eluted in less than 20 minutes. Mobile phasesare standard reverse-phase mobile phases: 0.1% formic acid in water and0.1% formic acid in acetonitrile. In these experiments backbone amidehydrogens that are exposed on the surface of IL-18 will incorporatedeuterons.

Next, triplicate labeling experiments were carried out. First, MOR9464antibody was immobilized on Protein G agarose beads (Thermo 22851) usingstandard techniques. Antibody beads were centrifuged to remove a PBSsolution. Then 200 μL of cold PBS (pH 7.4) and 6 μL (316 pmol) of IL-18was added to the immobilized MOR9464 antibody and incubated for 15 minat 4° C. After incubation, the complex was centrifuged and washed with200 μL PBS and centrifuged again. For deuterium exchange, 200 μL of83.6% deuterium PBS buffer was added to the antigen-antibody complex forincubation at 4° C. for 3 or 25 min. Deuterium buffer was then removed,and immediately, 125 μL quench buffer (as above) was added. Afterquenching, the flow-through was transferred into pre-chilled HPLC vialand analysed using the identical on line pepsin digestion/LC-MS analysisthat was in the control experiments. The backbone amides that arepresent in the antigen-antibody interface will incorporate fewerdeuterons at a given exchange time relative to the control experiments.By comparing the H/Dx patterns of the labeling and control experiments,the epitope is revealed as that area of the antigen that is protectedfrom on-exchange in the labelling experiments.

The results of this analysis, when conducted with MOR9464, revealedthree most significant regions of protection on IL-18. These were (withreference to SEQ ID NO:1) from amino acid 87 to 99, from amino acid 119to 137 and from amino acid 138 to 160. These regions are shown tocontain amino acids which are involved in the binding of the IL-18BP(FIG. 12).

9.11) X-Ray Structural Characterization of IL-18/Antibody Fragment

The general epitope identification provided by modelling and H/DxMSconfirmed that the antibodies and fragments thereof as described hereinrecognise a region on IL-18 which is also recognised by the IL-18BP(FIG. 12). In order to identify the relevant amino acids on IL-18 inthese regions, X-ray structure determination of an IL-18/antibodycomplex was carried out.

To prepare the antibody fragment, 13 mg of MOR9464_N30K at aconcentration of 18 mg/mL in 10 mM Histidine pH 5.0, were cleaved with1/300 (w/w) papain in 100 mM Tris-HCl pH 7.0, 10 mM DTT during 200 minat room temperature. The reaction was stopped with 50 μM of papaininhibitor E64. The Fab fragment was then purified over a protein Acolumn equilibrated with 20 mM sodium phosphate buffer pH 7.0.

Purification of the Fab complex with IL-18: A 1.33-fold excess of humanIL-18 (1.6 mg in PBS) was added to 3.2 mg of the MOR9464_N30K antibodyfragment (recovered from the Protein A flow-through). The IL-18 complexwith the MOR9464_N30K antibody fragment was concentrated byultrafiltration, loaded on a SPX-75 size-exclusion chromatography andeluted isocratically in 10 mM Tris-HCl pH 7.4, 25 mM NaCl.

Crystallization: The IL-18/MOR9464_N30K antibody fragment complex wasconcentrated by ultrafiltration to 11.8 mg/mL and crystallizationscreening was performed by vapor diffusion in sitting drops in 96-wellplates. The experiments were set up with a Phoenix robotic system andstored in a RockImager hotel at 19° C. Two crystal forms (form A andform B) were identified and characterized:

1) Crystal form A grew from 0.1M Lithium sulfate, 0.1M ADA pH 6.5, 12%PEG 4,000. (Cryo-protectant was a 1:1 mix of the reservoir solution with20% PEG 4,000, 30% glycerol)

2) Crystal form B grew from 59.5% 2-methyl-2,4-pentanediol (MPD), 15%glycerol, 85 mM HEPES pH 7.5. (No cryo-protectant needed).

X-ray data were collected at the Swiss Light Source, beamline X10SA,with a Pilatus pixel detector. All diffraction images were processedwith XDS (version Dec. 6, 2010), as implemented in APRV.

For crystal form A, 720 images of 0.25° oscillation each were recordedat a crystal-to-detector distance of 460 mm, using X-ray radiation of0.99999 Å wavelength.

For crystal form B, 720 images of 0.25° oscillation each were recordedat a crystal-to-detector distance of 430 mm, using X-ray radiation of0.99984 Å wavelength.

The structure of crystal form A was determined by molecular replacementwith the program Phaser, using PDB entry 3GBM.pdb and 2VXT.pdb asstarting models for the MOR9464_N30K antibody fragment and IL-18molecule, respectively. The variable and first constant domains of theantibody fragment in 3GBM.pdb were used as independent search models. Aclear solution for one IL-18/MOR9464_N30K antibody fragment complex perasymmetric unit was readily obtained. The structure of crystal form Bwas determined in the same way, but using the refined models derivedfrom crystal form A instead of the PDB entries.

Structure refinement: The structure was refined by multiple cycles ofelectron-density map inspection and model rebuilding in Coot 0.6.2followed by automated refinement using autoBUSTER (1.11.2/buster2.11.2). Intermolecular contacts were identified with NCONT and theburied surface was analyzed with AREAIMOL, both from the CCP4 programsuite (version 6.1.2). X-ray data collection and refinement statisticsare shown in Table 4 below.

TABLE 4 X-ray data collection and refinement statistics Crystal form ACrystal form B Data collection Space group P2₁ C2 Cell dimensions a, b,c (Å) 43.46, 85.86, 85.77 213.72, 41.81, 71.37 α, β, γ (°) 90.00, 94.29,90.00 90.00, 100.13, 90.00 Resolution (Å) 2.80 (2.87-2.80)* 2.70(2.77-2.70)* R_(sym) or R_(merge) 0.10 (0.454) 0.074 (0.487) I/σI 13.0(3.2) 12.0 (2.8) Completeness (%) 98.7 (98.2) 98.8 (98.6) Redundancy 3.4(3.5) 3.3 (3.4) Refinement Resolution (Å) 85.53-2.80 54.19-2.70 No.reflections 15,406 17,335 R_(work)/R_(free) 0.175/0.257 0.192/0.254 No.atoms Protein 4,447 4,445 Buffer component 10 (2 sulfate ions) 6 (1glycerol) Water 89 62 B-factors (Å²) Ab light-chain (L) 54.3 (V_(L):34.6; V_(C): 76.5) 50.3 Ab heavy-chain (H) 37.0 51.4 hIL-18 (I) 35.364.7 Water 30.4 51.6 R.m.s. deviations Bond lengths (Å)/ 0.010/1.31 0.010/1.28  angles (°)

The overall view of the structure of the complex of IL-18/MOR9464_N30Kfragment is shown in FIG. 13. Upon complex formation, 36 amino acids onIL-18 with reduced solvent accessibility are identified at the bindinginterface of IL-18 with the antibody fragment. These residues are Leu41,Glu42, Met87, Tyr88, Lys89, Asp90, Ser91, Gln92, Pro93, Arg94, Gly95,Met96, A1a97, Phe138, Gln139, Arg140, Ser141, Val142, Pro143, Gly144,His145, Asp146, Asn147, Met149, Gln150, Glu152, Ser153, Ser154, Glu157,Gly158, Phe160, Glu177, Asp178, Glu179, Leu180 and Gly181.

Further characterization of the relevant contacts made in the complexinterface identified that Arg140 and Glu152 are the amino acids whichare likely to contribute the most to this particular complex formation.Both residues are located at the centre of the binding interface, andcontribute to a large number of intermolecular contacts (21 and 18,respectively, using a cut-off distance of 4.0 Å). Arg140 interacts withboth L-CDR3 (Tyr94L) and H-CDR3 (Tyr101H, His102H) residues. Glu152forms a strong (buried) salt-bridge interaction with L-CDR2 Arg51L andaccepts a H-bond from L-CDR3 Tyr94L. All residues in the antibody chainsare numbered sequentially, the letter L or H following the residuenumber indicate a residue in the light chain or heavy chain,respectively.

The three dimensional structure of IL-18/MOR9464_N30K antibody fragmentcomplex allowed some further investigations on the differentcross-reactivity between the human IL-18 and the cynomolgus IL-18.MOR9464_N30K is 10-fold weaker against cynomolgus IL-18 (20 pM) comparedto 2 pM against human IL-18 (Table 2). Cynomolgus IL-18 differs fromhuman IL-18 in 6 positions (human vs cynomolgus): V471; S86N; T99A;K115R; F170Y and E177K (FIG. 14(A). Of these positions only glutamicacid 177 (Glu177; E177) is at the IL-18/antibody fragment complexinterface (FIG. 14(B)). Thus, it is arguable that K177 in thecynomolgous monkey sequence of cynomolgus IL-18 causes a 10-fold drop inaffinity to MOR9464_N30K as indicated in Table 2A column 1, where theK_(D) (SET, pM) for MOR9464_N30K for human IL-18 is 2±1 pM whereas forcynomolgus IL-18 is 20±10 pM.

In order to identify with what part of the IL-18Rα MOR9464_N30K antibodyfragment competes, a superposition of the IL-18/MOR9464_N30K antibodyfragment complex with the IL-18/IL-18Rα complex was performed. In brief,the structures of IL-18 in the IL-18/IL-18Rα complex and IL-18 in theIL-18/MOR9464_N30K complex were superposed by using the align command inPyMol (The PyMol Molecular Graphic system, version 1.2r3pre,Schroedinger LLC).

As shown in FIG. 15, it appears that MOR9464_N30K antibody fragmentcompetes with the Ig-domain D3 of IL-18Rα for binding IL-18.

Similarly, the structure of the IL-18/MOR9464_N30K antibody fragmentcomplex was superimposed onto the structure of the human IL-18 incomplex with poxvirus IL-18BP (coordinate file used 3F62.pdb) using the“align” command in PyMOL (The PyMOL Molecular Graphic system, version1.5.0., Schrödinger LLC).

As shown in FIG. 16, the poxvirus IL-18BP clashes with the heavy chainvariable domain of the MOR9464_N30K antibody fragment and competespredominantly for binding amino acids residues Met87 to Met96 on IL-18.

These findings further confirm the biochemical and epitope binning datashowed herein and show that the binding molecules as described herein,in particular the antibodies and the fragments thereof, do not bind theIL-18/IL-18BP complex.

Finally, comparison with the prior art murine antibody 125-2H wasperformed. The analysis was carried out with the structure of 125-2Hantibody fragment as found in the 2VXT.pdb file. In brief, the structureof IL-18 in complex with MOR9464_N30K antibody fragment was superimposedonto IL-18 in complex with the 125-2H Fab using the “align” command inPyMOL (The PyMOL Molecular Graphic system, version 1.5.0., SchrödingerLLC).

As shown in FIG. 17, MOR9464_N30K antibody fragment (left side of thesuperimposition) and 125-2H antibody fragment (right side of thesuperimposition) overlap in recognising IL-18. However, 125-2H antibodyfragment does not bind the epitope recognised by the IL-18BP as shown inFIG. 18 where the 125-2H/IL-18 complex was superposed with the poxvirusIL-18BP/IL complex. These data further confirm, both the prior artliterature with regard to 125-2H (Argiriadi M. A et al (2009) J. Biol.Chem. 284:24478) and also the biochemical and epitope binning datashowed herein that murine antibody 125-2H binds the IL-18/IL-18BPcomplex.

Finally, as shown in FIG. 19, Lysine in position 30 in the heavy chainof MOR9464_N30K appears to form electrostatic/polar interactions withAsp 146 and Asn 147 of human IL-18 indicating that lysine 30 is involvedin the formation of the antibody-antigen complex. It is, therefore,apparent that an exocellular modification as described in section 9.2herein such as asparagine deamidation, may have contributed to the lossof potency over time of MOR9464. Replacement of asparagine 30 with alysine in MOR9464_N30K appears to provide MOR9464_N30K with increasedstability.

Sequence Correlation Table SEQ ID NO: Identity 1 Human IL-18 2Cynomolgus IL-18 3H-CDR1 of MOR8775; MOR9464; MOR9441; MOR10222; MOR10579; MOR9464_N30K;MOR10222_N30S_M54I; MOR9465; MOR9466; 4 H-CDR2 of MOR8775; 5H-CDR3 of MOR8775; MOR9464; MOR9441; MOR10222; MOR10579; MOR9464_N30K;MOR10222_N30S_M54I; MOR9465; MOR9466; 6L-CDR1 of MOR8775; MOR9464; MOR9441; MOR10222; MOR10579; MOR9464_N30K;MOR10222_N30S_M54I; MOR9465; MOR9466; 7L-CDR2 of MOR8775; MOR9464; MOR9441; MOR10222; MOR10579; MOR9464_N30K;MOR10222_N30S_M54I; MOR9465; MOR9466; 8L-CDR3 of MOR8775; MOR9464; MOR9441; MOR10222; MOR10579; MOR9464_N30K;MOR10222_N30S_M54I; MOR9465; MOR9466; 9H-CDR2 of MOR9464; MOR10222; MOR9464_N30K; 10H-CDR2 of MOR9441; MOR10579; 11 H-CDR2 of MOR9465 12 H-CDR2 of MOR946613 H-CDR2 of MOR10222_N30S_M54I; 14 VH of MOR9464_N30K 15Polynucleotide VH of MOR9464_N30K 16VL of MOR9464_N30K; MOR9464; MOR8775; MOR9465; MOR9466; MOR9441 17Polynucleotide VL of MOR9464_N30K 18 VH of MOR10222_N30S_M54I 19Polynucleotide VH of MOR10222_N30S_M54I 20VL of MOR10222_N30S_M54I; MOR10579; MOR10222 21Polynucleotide VL of MOR10222_N30S_M54I 22 VH MOR8775; 23Polynucleotide VH MOR8775; 24 Polynucleotide VL MOR8775; 25 VH MOR9441;26 Polynucleotide VH MOR9441; 27 Polynucleotide VL MOR9441; 28VH MOR9464; 29 Polynucleotide VH MOR9464; 30 Polynucleotide VL MOR9464;31 VH MOR9465; 32 Polynucleotide VH MOR9465; 33Polynucleotide VL MOR9465; 34 VH MOR9466; 35 Polynucleotide VH MOR9466;36 Polynucleotide VL MOR9466; 37 VH MOR10579; 38Polynucleotide VH MOR10579; 39 Polynucleotide VL MOR10579; 40VH MOR10222; 41 Polynucleotide VH MOR10222; 42Polynucleotide VL MOR10222; 43 Heavy chain of MOR9464_N30K 44Polynucleotide Heavy chain of MOR9464_N30K 45Light chain of MOR9464_N30K; MOR9464; MOR8775; MOR9441 46Polynucleotide Light chain of MOR9464_N30K 47 Heavy Chain MOR9464; 48Polynucleotide Heavy Chain MOR9464; 49Polynucleotide Light Chain MOR9464; 50 Heavy Chain MOR9441; 51Polynucleotide Heavy Chain MOR9441; 52Polynucleotide Light Chain MOR9441; 53 Heavy Chain MOR10222; 54Polynucleotide Heavy Chain MOR10222; 55Polynucleotide Light Chain MOR10222; 56 Heavy Chain MOR8775; 57Polynucleotide Heavy Chain MOR8775; 58Polynucleotide Light Chain MOR8775; 59 (Chothia) H-CDR1 MOR9464; 60(Chothia) H-CDR2 MOR9464; MOR9464_N30K 61(Chothia) H-CDR3 MOR9464; MOR9464_N30K; MOR10222_N30S_M54I 62(Chothia) L-CDR1 MOR9464; MOR9464_N30K; MOR10222_N30S_M54I 63(Chothia) L-CDR2 MOR9464; MOR9464_N30K; MOR10222_N30S_M54I 64(Chothia) L-CDR3 MOR9464; MOR9464_N30K; MOR10222_N30S_M54I 65(Chothia) H-CDR1 MOR9464_N30K; 66 (Chothia) H-CDR1 MOR10222_N30S_M54I 67(Chothia) H-CDR2 MOR10222_N30K_M54I; 68 (Chothia) H-CDR1 MOR13363; 69(Chothia) H-CDR2 MOR13363; 70 (Chothia) H-CDR3 MOR13363; 71(Chothia) L-CDR1 MOR13363; 72 (Chothia) L-CDR2 MOR13363; 73(Chothia) L-CDR3 MOR13363; 74H-CDR1 of MOR8776; MOR10497; MOR10501; MOR10502; 75 H-CDR2 of MOR8776;76 H-CDR2 of MOR10501 77 H-CDR2 of MOR1010502; 78 H-CDR2 of MOR10497; 79H-CDR3 of MOR8776; MOR10497; MOR10501; MOR10502; 80L-CDR1 of MOR8776; MOR10497; MOR10501; MOR10502; 81L-CDR2 of MOR8776; MOR10497; MOR10501; MOR10502; 82L-CDR3 of MOR8776; MOR10497; MOR10501; MOR10502; 83 VH MOR8776; 84Polynucleotide VH MOR8776; 85 VL MOR8776; MOR10497; MOR10501; MOR1050286 Polynucleotide VL MOR8776; 87 VH MOR10497; 88Polynucleotide VH MOR10497; 89 Polynucleotide VL MOR10497; 90VH MOR10501; 91 Polynucleotide VH MOR10501; 92Polynucleotide VL MOR10501; 93 VH MOR10502; 94Polynucleotide VH MOR10502; 95 Polynucleotide VL MOR10502; 96Heavy Chain of MOR8776; 97 Polynucleotide Heavy Chain of MOR8776; 98Light Chain of MOR8776; MOR10497 99Polynucleotide Light Chain of MOR8776; 100 Heavy Chain MOR10579; 101Polynucleotide Heavy Chain MOR10579; 102Polynucleotide Light Chain MOR10579; 103 Heavy Chain MOR10497; 104Polynucleotide Heavy Chain MOR10497; 105Polynucleotide Light Chain MOR10497; 106 H-CDR1 of MOR13363; MOR13361107 H-CDR2 of MOR13363 108 H-CDR3 of MOR13363; MOR13361 109L-CDR1 of MOR13363; MOR13361 110 L-CDR2 of MOR13363; MOR13361 111L-CDR3 of MOR13363 112 VH of MOR13363 113 Polynucleotide VH of MOR13363114 VL of MOR13363 115 Polynucleotide VL of MOR13363 116Heavy Chain of MOR13363 117 Polynucleotide Heavy Chain of MOR13363 118Light Chain of MOR13363 119 Polynucleotide Light Chain of MOR13363 120H-CDR1 MOR13341; MOR13342; MOR13347 121H-CDR2 MOR13341; MOR13342; MOR13347 122 H-CDR2 MOR13361 123H-CDR3 MOR13341; MOR13342; MOR13347 124L-CDR1 MOR13341; MOR13342; MOR13347 125L-CDR2 MOR13341; MOR13342; MOR13347 126 L-CDR3 MOR13361 127L-CDR3 MOR13341 128 L-CDR3 MOR13342 129 L-CDR3 MOR13347 130VH MOR13341; MOR13342; MOR13347 131 Polynucleotide VH MOR13341 132VL MOR13341 133 Polynucleotide VL MOR13341 134Heavy chain MOR13341; MOR13342; MOR13347 135Polynucleotide Heavy chain MOR13341 136 Light chain MOR13341 137Polynucleotide Light chain MOR13341 138 VH MOR13361 139Polynucleotide VH MOR13361 140 VL MOR13361 141Polynucleotide VL MOR13361 142 Heavy chain MOR13361 143Polynucleotide Heavy chain MOR13361 144 Light chain MOR13361 145Polynucleotide Light chain MOR13361 146 Polynucleotide VH MOR13342 147VL MOR13342 148 Polynucleotide VL MOR13342 149Polynucleotide Heavy chain MOR13342 150 Light chain MOR13342 151Polynucleotide Light chain MOR13342 152 Polynucleotide VH MOR13347 153VL MOR13347 154 Polynucleotide VL MOR13347 155Polynucleotide Heavy chain MOR13347 156 Light chain MOR13347 157Polynucleotide Light chain MOR13347 158Heavy Chain of MOR10222_N30S_M54I 159Polynucleotide Heavy Chain of MOR10222_N30S_M54I 160Light Chain of MOR10222_N30S_M54I; MOR10222; MOR10579 161Polynucleotide Light Chain of MOR10222_N30S_M54I 162(Chothia) H-CDR1 MOR10497 163 (Chothia) H-CDR2 MOR10497 164(Chothia) H-CDR3 MOR10497 165 (Chothia) L-CDR1 MOR10497 166(Chothia) L-CDR2 MOR10497 167 (Chothia) L-CDR3 MOR10497 168Heavy chain MOR10501 169 Polynucleotide Heavy chain MOR10501 170Light chain MOR10501 171 Polynucleotide Light chain MOR10501 172Heavy chain MOR10502 173 Polynucleotide Heavy chain MOR10502 174Light chain MOR10502 175 Polynucleotide Light chain MOR10502 176Heavy chain MOR9465 177 Polynucleotide Heavy chain MOR9465 178Light chain MOR9465 179 Polynucleotide Light chain MOR9465 180Heavy chain MOR9466 181 Polynucleotide Heavy chain MOR9466 182Light chain MOR9466 183 Polynucleotide Light chain MOR9466 184(Chothia) H-CDR1 MOR8776 185 (Chothia) H-CDR2 MOR8776 186(Chothia) H-CDR3 MOR8776 187 (Chothia) L-CDR1 MOR8776 188(Chothia) L-CDR2 MOR8776 189 (Chothia) L-CDR3 MOR8776 190(Chothia) H-CDR1 MOR10501 191 (Chothia) H-CDR2 MOR10501 192(Chothia) H-CDR3 MOR10501 193 (Chothia) L-CDR1 MOR10501 194(Chothia) L-CDR2 MOR10501 195 (Chothia) L-CDR3 MOR10501 196H-CDR2 MOR10502 197 (Chothia) H-CDR1 MOR10502 198(Chothia) H-CDR2 MOR10502 199 (Chothia) H-CDR3 MOR10502 200(Chothia) L-CDR1 MOR10502 201 (Chothia) L-CDR2 MOR10502 202(Chothia) L-CDR3 MOR10502 203 (Chothia) H-CDR1 MOR8775 204(Chothia) H-CDR2 MOR8775 205 (Chothia) H-CDR3 MOR8775 206(Chothia) L-CDR1 MOR8775 207 (Chothia) L-CDR2 MOR8775 208(Chothia) L-CDR3 MOR8775 209 (Chothia) H-CDR1 MOR9441 210(Chothia) H-CDR2 MOR9441 211 (Chothia) H-CDR3 MOR9441 212(Chothia) L-CDR1 MOR9441 213 (Chothia) L-CDR2 MOR9441 214(Chothia) L-CDR3 MOR9441 215 (Chothia) H-CDR1 MOR9465 216(Chothia) H-CDR2 MOR9465 217 (Chothia) H-CDR3 MOR9465 218(Chothia) L-CDR1 MOR9465 219 (Chothia) L-CDR2 MOR9465 220(Chothia) L-CDR3 MOR9465 221 (Chothia) H-CDR1 MOR9466 222(Chothia) H-CDR2 MOR9466 223 (Chothia) H-CDR3 MOR9466 224(Chothia) L-CDR1 MOR9466 225 (Chothia) L-CDR2 MOR9466 226(Chothia) L-CDR3 MOR9466 227 (Chothia) H-CDR1 MOR10579 228(Chothia) H-CDR2 MOR10579 229 (Chothia) H-CDR3 MOR10579 230(Chothia) L-CDR1 MOR10579 231 (Chothia) L-CDR2 MOR10579 232(Chothia) L-CDR3 MOR10579 233 (Chothia) H-CDR1 MOR10222 234(Chothia) H-CDR2 MOR10222 235 (Chothia) H-CDR3 MOR10222 236(Chothia) L-CDR1 MOR10222 237 (Chothia) L-CDR2 MOR10222 238(Chothia) L-CDR3 MOR10222 239 H-CDR2 MOR10222_N30S_M54I 240(Chothia) H-CDR1 MOR13341 241 (Chothia) H-CDR2 MOR13341 242(Chothia) H-CDR3 MOR13341 243 (Chothia) L-CDR1 MOR13341 244(Chothia) L-CDR2 MOR13341 245 (Chothia) L-CDR3 MOR13341 246(Chothia) H-CDR1 MOR13342 247 (Chothia) H-CDR2 MOR13342 248(Chothia) H-CDR3 MOR13342 249 (Chothia) L-CDR1 MOR13342 250(Chothia) L-CDR2 MOR13342 251 (Chothia) L-CDR3 MOR13342 252(Chothia) H-CDR1 MOR13347 253 (Chothia) H-CDR2 MOR13347 254(Chothia) H-CDR3 MOR13347 255 (Chothia) L-CDR1 MOR13347 256(Chothia) L-CDR2 MOR13347 257 (Chothia) L-CDR3 MOR13347 258(Chothia) H-CDR1 MOR13361 259 (Chothia) H-CDR2 MOR13361 260(Chothia) H-CDR3 MOR13361 261 (Chothia) L-CDR1 MOR13361 262(Chothia) L-CDR2 MOR13361 263 (Chothia) L-CDR3 MOR13361 Sequence ListingSEQ ID NO: 1MAAEPVEDNCINFVAMKFIDNTLYFIAEDDENLESDYFGKLESKLSVIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFIISMYKDSQPRGMAVTISVKCEKISTLSCENKIISFKEMNPPDNIKDTKSDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLFKLILKKEDELGDRSIMFTVQNED SEQ ID NO: 2MAAEPAEDNCINFVAMKPIDSTLYFIAEDDENLESDYFGKLESKLSIIRNLNDQVLFIDQGNRPLFEDMTDSDCRDNAPRTIFIINMYKDSQPRGMAVAISVKCEKISTLSCENRIISFKEMNPPDNIKDTKSDIIFFQRSVPGHDNKMQFESSSYEGYFLACEKERDLYKLILKKKDELGDRSIMFTVQNED SEQ ID NO: 3 SYAIS SEQ ID NO: 4GIIPIYGTANYAQKFQG SEQ ID NO: 5 AAYHPLVFDN SEQ ID NO: 6 SGSSSNIGNHYVNSEQ ID NO: 7 RNNHRPS SEQ ID NO: 8 QSWDYSGFSTV SEQ ID NO: 9NIIPMTGQTYYAQKFQG SEQ ID NO: 10 WINPFYIGETFYAQKFQG SEQ ID NO: 11NIIPHYGFAYYAQKFQG SEQ ID NO: 12 NIIPYSGFAYYAQKFQG SEQ ID NO: 13NIIPITGQTYYAQKFQG SEQ ID NO: 14EVQLVQSGAEVKKPGSSVKVSCKASGGTFKSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 15GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAAGTCCTACGCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCAATATTATCCCTATGACCGGTCAGACCTACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGAGCCGCCTATCACCCCCTGGTGTTCGATAACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 16DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVL SEQ ID NO: 17GATATCGTCCTGACTCAGCCCCCTAGCGTCAGCGGCGCTCCCGGTCAGAGAGTGACTATTAGCTGTAGCGGCTCTAGCTCTAATATCGGTAATCACTACGTGAACTGGTATCAGCAGCTGCCCGGCACCGCCCCTAAGCTGCTGATCTATAGAAACAATCACCGGCCTAGCGGCGTGCCCGATAGGTTTAGCGGATCTAAGTCAGGCACTAGCGCTAGTCTGGCTATCACCGGACTGCAGTCAGAGGACGAGGCCGACTACTACTGTCAGTCCTGGGACTATAGCGGCTTTAGCACCGTGTTCGGCGGAGGCACTAAGCTGACCGTGCTG SEQ ID NO: 18QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGNIIPITGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 19CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCTCTAGCTACGCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCAATATTATCCCTATCACCGGTCAGACCTACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGAGCCGCCTATCACCCCCTGGTGTTCGATAACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 20QSVLTQPPSASGTPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVL SEQ ID NO: 21CAGTCAGTCCTGACTCAGCCCCCTAGCGCTAGTGGCACCCCTGGTCAGAGAGTGACTATTAGCTGTAGCGGCTCTAGCTCTAATATCGGTAATCACTACGTGAACTGGTATCAGCAGCTGCCCGGCACCGCCCCTAAGCTGCTGATCTATAGAAACAATCACCGGCCTAGCGGCGTGCCCGATAGGTTTAGCGGATCTAAGTCAGGGACTAGCGCTAGTCTGGCTATTAGCGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGTCAGTCCTGGGACTATAGCGGCTTTAGCACCGTGTTCGGCGGAGGCACTAAGCTGACCGTGCTG SEQ ID NO: 22QVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGGIIPIYGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 23CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCATTCCGATTTATGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCA SEQ ID NO: 24GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 25EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGWINPFYIGETFYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 26GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCTGGATCAACCCTTTCTACATCGGCGAGACATTCTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCC SEQ ID NO: 27GATATCGTGCTGACCCAGCCTCCTTCTGTGTCTGGCGCCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCTGCCTCCCTGGCCATCACCGGCCTGCAGTCCGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCAACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTG SEQ ID NO: 28EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 29GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCAACATCATCCCTATGACCGGCCAGACCTACTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCC SEQ ID NO: 30GACATCGTGCTGACACAGCCTCCCTCTGTGTCTGGCGCCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCTGCCTCCCTGGCCATCACCGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTG SEQ ID NO: 31EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPHYGFAYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 32GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATTATTCCTCATTATGGTTTTGCTTATTATGCTCAGAAGTTTCAGGGTCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCA SEQ ID NO: 33GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 34EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPYSGFAYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 35GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATTATTCCTTATTCTGGTTTTGCTTATTATGCTCAGAAGTTTCAGGGTCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCA SEQ ID NO: 36GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 37EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGWINPFYIGETFYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 38GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCTGGATCAACCCTTTCTACATCGGCGAGACATTCTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCC SEQ ID NO: 39CAGTCCGTGCTGACCCAGCCTCCTTCTGCCTCTGGCACCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCCGCCTCCCTGGCCATCTCTGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTG SEQ ID NO: 40EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSS SEQ ID NO: 41GAGGTGCAGCTGGTGCAGTCTGGCGCCGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCCATCTCTTGGGTGCGCCAGGCTCCTGGACAGGGCCTGGAGTGGATGGGCAACATCATCCCTATGACCGGCCAGACCTACTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCC SEQ ID NO: 42CAGTCCGTGCTGACCCAGCCTCCTTCTGCCTCTGGCACCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCCGCCTCCCTGGCCATCTCTGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTG SEQ ID NO: 43EVQLVQSGAEVKKPGSSVKVSCKASGGTFKSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 44GAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAAGTCCTACGCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCAATATTATCCCTATGACCGGTCAGACCTACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGAGCCGCCTATCACCCCCTGGTGTTCGATAACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 45DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 46GATATCGTCCTGACTCAGCCCCCTAGCGTCAGCGGCGCTCCCGGTCAGAGAGTGACTATTAGCTGTAGCGGCTCTAGCTCTAATATCGGTAATCACTACGTGAACTGGTATCAGCAGCTGCCCGGCACCGCCCCTAAGCTGCTGATCTATAGAAACAATCACCGGCCTAGCGGCGTGCCCGATAGGTTTAGCGGATCTAAGTCAGGCACTAGCGCTAGTCTGGCTATCACCGGACTGCAGTCAGAGGACGAGGCCGACTACTACTGTCAGTCCTGGGACTATAGCGGCTTTAGCACCGTGTTCGGCGGAGGCACTAAGCTGACCGTGCTGGGTCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 47EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 48GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCAACATCATCCCTATGACCGGCCAGACCTACTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCCGCTAGCACCAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 49GACATCGTGCTGACACAGCCTCCCTCTGTGTCTGGCGCCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCTGCCTCCCTGGCCATCACCGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTGGGACAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 50EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGWINPFYIGETFYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 51GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCTGGATCAACCCTTTCTACATCGGCGAGACATTCTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCCGCTAGCACCAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 52GATATCGTGCTGACCCAGCCTCCTTCTGTGTCTGGCGCCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCTGCCTCCCTGGCCATCACCGGCCTGCAGTCCGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCAACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTGGGACAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 53EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPMTGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 54GAGGTGCAGCTGGTGCAGTCTGGCGCCGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCCATCTCTTGGGTGCGCCAGGCTCCTGGACAGGGCCTGGAGTGGATGGGCAACATCATCCCTATGACCGGCCAGACCTACTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCCGCTAGCACCAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 55CAGTCCGTGCTGACCCAGCCTCCTTCTGCCTCTGGCACCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCCGCCTCCCTGGCCATCTCTGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTGGGACAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 56QVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGGIIPIYGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 57CAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCGGTATCATTCCGATTTATGGCACTGCGAATTACGCGCAGAAGTTTCAGGGCCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAASEQ ID NO: 58GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 59 GGTFNSYSEQ ID NO: 60 IPMTGQ SEQ ID NO: 61 AAYHPLVFDN SEQ ID NO: 62 SSSNIGNHYSEQ ID NO: 63 RNN SEQ ID NO: 64 WDYSGFST SEQ ID NO: 65 GGTFKSYSEQ ID NO: 66 GGTFSSY SEQ ID NO: 67 IPITGQ SEQ ID NO: 68 GFTFSSYSEQ ID NO: 69 SGEGSN SEQ ID NO: 70 VMIGYGFDY SEQ ID NO: 71 SQSIFNYSEQ ID NO: 72 DSS SEQ ID NO: 73 YSGFLF SEQ ID NO: 74 TGSYYWNSEQ ID NO: 75 EINHMGITYYNPSLKG SEQ ID NO: 76 EIWHSGPTFYNPSLKSSEQ ID NO: 77 EIHGHGFTFYNPSLKS SEQ ID NO: 78 EIQSPGYTFYNPSLKSSEQ ID NO: 79 TTRYWMSHILAYGMDY SEQ ID NO: 80 SGSSSNIGNHYVS SEQ ID NO: 81ANTKRPS SEQ ID NO: 82 SSYDGSQSIV SEQ ID NO: 83QVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEINHMGITYYNPSLKGRVTISVDTSKNQFSLKLSSVTAEDTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSS SEQ ID NO: 84CAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATCAATCATATGGGCATTACCTATTATAATCCGAGCCTGAAAGGCCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGAAGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCASEQ ID NO: 85DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVSWYQQLPGTAPKLLIYANTKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCSSYDGSUSIVEGGGTKLTVL SEQ ID NO: 86GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 87EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIQSPGYTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSS SEQ ID NO: 88GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTCAGTCTCCTGGTTATACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCASEQ ID NO: 89GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 90EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIWHSGPTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSS SEQ ID NO: 91GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTTGGCATTCTGGTCCTACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCASEQ ID NO: 92GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 93EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIHGHGFTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSS SEQ ID NO: 94GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTCATGGTCATGGTTTTACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCASEQ ID NO: 95GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTA SEQ ID NO: 96QVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEINHMGITYYNPSLKGRVTISVDTSKNQFSLKLSSVTAEDTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 97CAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATCAATCATATGGGCATTACCTATTATAATCCGAGCCTGAAAGGCCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGAAGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA SEQ ID NO: 98DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVSWYQQLPGTAPKLLIYANTKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCSSYDGSQSIVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 99GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 100EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGWINPFYIGETFYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 101GAGGTGCAGCTGGTGCAGTCTGGCGCTGAGGTGAAGAAGCCTGGCTCCTCCGTCAAGGTGTCCTGCAAGGCCTCCGGCGGCACCTTCAACTCCTACGCTATCTCTTGGGTGCGCCAGGCTCCCGGACAGGGCCTGGAGTGGATGGGCTGGATCAACCCTTTCTACATCGGCGAGACATTCTACGCCCAGAAGTTCCAGGGCAGAGTCACCATCACCGCCGACGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCAGAGGACACCGCCGTGTACTACTGCGCCAGGGCCGCCTACCACCCTCTGGTGTTCGACAACTGGGGCCAGGGCACCCTGGTGACCGTGTCCTCCGCTAGCACCAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 102CAGTCCGTGCTGACCCAGCCTCCTTCTGCCTCTGGCACCCCTGGCCAGAGAGTGACCATCTCCTGCTCTGGCTCCTCCTCCAATATCGGCAACCACTACGTGAACTGGTATCAGCAGCTGCCCGGAACCGCCCCTAAGCTGCTGATCTACCGGAACAACCACCGGCCTTCCGGCGTGCCCGACCGGTTCTCCGGCTCCAAGTCTGGCACCTCCGCCTCCCTGGCCATCTCTGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGCCAGTCCTGGGACTACTCCGGCTTCTCCACCGTGTTCGGCGGAGGCACCAAGCTGACCGTGCTGGGACAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 103EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIQSPGYTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 104GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTCAGTCTCCTGGTTATACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA SEQ ID NO: 105GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 106 SYAIHSEQ ID NO: 107 VISGEGSNTYYADSVKG SEQ ID NO: 108 VMIGYGFDY SEQ ID NO: 109RASQSIFNYLN SEQ ID NO: 110 DSSTLQS SEQ ID NO: 111 LQYSGFLFTSEQ ID NO: 112QVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAIHWVRQAPGKGLEWVSVISGEGSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVMIGYGFDYWGQGTLVTVSS SEQ ID NO: 113CAGGTGCAGCTGCTGGAATCAGGCGGCGGACTGGTGCAGCCTGGCGGTAGCCTGAGACTGAGCTGCGCTGCTAGTGGCTTCACCTTCTCTAGCTACGCTATTCACTGGGTCAGACAGGCCCCTGGTAAAGGCCTGGAGTGGGTGTCAGTGATTAGCGGCGAGGGCTCTAACACCTACTACGCCGATAGCGTGAAGGGCCGGTTCACTATCTCTAGGGATAACTCTAAGAACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAGGACACCGCCGTCTACTACTGCGCTAGAGTGATGATCGGCTACGGCTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 114DIQMTQSPSSLSASVGDRVTITCRASQSIFNYLNWYQQKPGKAPKLLIYDSSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYSGFLFTFGQGTKVEIK SEQ ID NO: 115GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTTTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACGACTCTAGCACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGCCTGCAGTATAGCGGCTTCCTGTTCACCTTCGGTCAGGGCACTAAGGTCGAGATTAAG SEQ ID NO: 116QVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAIHWVRQAPGKGLEWVSVISGEGSNTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVMIGYGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 117CAGGTGCAGCTGCTGGAATCAGGCGGCGGACTGGTGCAGCCTGGCGGTAGCCTGAGACTGAGCTGCGCTGCTAGTGGCTTCACCTTCTCTAGCTACGCTATTCACTGGGTCAGACAGGCCCCTGGTAAAGGCCTGGAGTGGGTGTCAGTGATTAGCGGCGAGGGCTCTAACACCTACTACGCCGATAGCGTGAAGGGCCGGTTCACTATCTCTAGGGATAACTCTAAGAACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAGGACACCGCCGTCTACTACTGCGCTAGAGTGATGATCGGCTACGGCTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 118DIQMTQSPSSLSASVGDRVTITCRASQSIFNYLNWYQQKPGKAPKLLIYDSSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYSGFLFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 119GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTTTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACGACTCTAGCACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGCCTGCAGTATAGCGGCTTCCTGTTCACCTTCGGTCAGGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 120 TFSISSEQ ID NO: 121 GIIPIFGTANYAQKFQG SEQ ID NO: 122 TIQSSGENKFYADSVKGSEQ ID NO: 123 GGYGGYYYFDY SEQ ID NO: 124 RASQSISNRLN SEQ ID NO: 125KGSTLQS SEQ ID NO: 126 HQYSGLLFT SEQ ID NO: 127 QQHKVWLTT SEQ ID NO: 128QQHYVWSTT SEQ ID NO: 129 QQHYQWLTT SEQ ID NO: 130QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTFSISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGGYGGYYYFDYWGQGTLVTVSS SEQ ID NO: 131CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAGCACCTTCTCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCGGAATTATCCCTATCTTCGGCACCGCTAACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGGGGCGGCTACGGCGGCTATTACTACTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCSEQ ID NO: 132DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHKVWLTTFGQGTKVEIK SEQ ID NO: 133GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTCTAATAGGCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTATAAGGGCTCTACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCAGCAGCACAAAGTGTGGCTGACTACCTTCGGTCAGGGCACTAAGGTCGAGATTAAG SEQ ID NO: 134QVQLVQSGAEVKKPGSSVKVSCKASGGTFSTFSISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGGYGGYYYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 135CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAGCACCTTCTCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCGGAATTATCCCTATCTTCGGCACCGCTAACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGGGGCGGCTACGGCGGCTATTACTACTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 136DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHKVWLTTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 137GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTCTAATAGGCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTATAAGGGCTCTACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCAGCAGCACAAAGTGTGGCTGACTACCTTCGGTCAGGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 138QVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAIHWVRQAPGKGLEWVSTIQSSGENKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVMIGYGFDYWGQGTLVTVSS SEQ ID NO: 139CAGGTGCAGCTGCTGGAATCAGGCGGCGGACTGGTGCAGCCTGGCGGTAGCCTGAGACTGAGCTGCGCTGCTAGTGGCTTCACCTTCTCTAGCTACGCTATTCACTGGGTCAGACAGGCCCCTGGTAAAGGCCTGGAGTGGGTCAGCACTATTCAGTCTAGCGGCGAGAACAAGTTCTACGCCGATAGCGTGAAGGGCCGGTTCACTATCTCTAGGGATAACTCTAAGAACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAGGACACCGCCGTCTACTACTGCGCTAGAGTGATGATCGGCTACGGCTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGC SEQ ID NO: 140DIQMTQSPSSLSASVGDRVTITCRASQSIFNYLNWYQQKPGKAPKLLIYDSSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYSGLLFTFGQGTKVEIK SEQ ID NO: 141GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTTTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACGACTCTAGCACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCACCAGTATAGCGGCCTGCTGTTCACCTTCGGTCAGGGCACTAAGGTCGAGATTAAG SEQ ID NO: 142QVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAIHWVRQAPGKGLEWVSTIQSSGENKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVMIGYGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 143CAGGTGCAGCTGCTGGAATCAGGCGGCGGACTGGTGCAGCCTGGCGGTAGCCTGAGACTGAGCTGCGCTGCTAGTGGCTTCACCTTCTCTAGCTACGCTATTCACTGGGTCAGACAGGCCCCTGGTAAAGGCCTGGAGTGGGTCAGCACTATTCAGTCTAGCGGCGAGAACAAGTTCTACGCCGATAGCGTGAAGGGCCGGTTCACTATCTCTAGGGATAACTCTAAGAACACCCTGTACCTGCAGATGAATAGCCTGAGAGCCGAGGACACCGCCGTCTACTACTGCGCTAGAGTGATGATCGGCTACGGCTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 144DIQMTQSPSSLSASVGDRVTITCRASQSIFNYLNWYQQKPGKAPKLLIYDSSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQYSGLLFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 145GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTTTAACTACCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTACGACTCTAGCACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCACCAGTATAGCGGCCTGCTGTTCACCTTCGGTCAGGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 146CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAGCACCTTCTCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCGGAATTATCCCTATCTTCGGCACCGCTAACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGGGGCGGCTACGGCGGCTATTACTACTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCSEQ ID NO: 147DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYVWSTTFGQGTKVEIK SEQ ID NO: 148GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTCTAATAGGCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTATAAGGGCTCTACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCAGCAGCACTACGTGTGGTCTACTACCTTCGGTCAGGGCACTAAGGTCGAGATTAAG SEQ ID NO: 149CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCAGCACCTTCTCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCGGAATTATCCCTATCTTCGGCACCGCTAACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGGGGCGGCTACGGCGGCTATTACTACTTCGACTACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 150DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYVWSTTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 151GATATTCAGATGACTCAGTCACCTAGTAGCCTGAGCGCTAGTGTGGGCGATAGAGTGACTATCACCTGTAGAGCCTCTCAGTCTATCTCTAATAGGCTGAACTGGTATCAGCAGAAGCCCGGTAAAGCCCCTAAGCTGCTGATCTATAAGGGCTCTACCCTGCAGTCAGGCGTGCCCTCTAGGTTTAGCGGTAGCGGTAGTGGCACCGACTTCACCCTGACTATCTCTAGCCTGCAGCCCGAGGACTTCGCTACCTACTACTGTCAGCAGCACTACGTGTGGTCTACTACCTTCGGTCAGGGCACTAAGGTCGAGATTAAGCGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCATAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACAGGGGCGAGTGC SEQ ID NO: 152CAGGTGCAATTGGTGCAGAGCGGTGCCGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTTAGCTGCAAAGCATCCGGAGGGACGTTTTCTACTTTCTCTATCTCTTGGGTGCGCCAGGCCCCGGGCCAGGGCCTCGAGTGGATGGGCGGTATCATCCCGATCTTCGGCACTGCGAACTACGCCCAGAAATTTCAGGGCCGGGTGACCATTACCGCCGATGAAAGCACCAGCACCGCCTATATGGAACTGAGCAGCCTGCGCAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGGTGGTTACGGTGGTTACTACTACTTCGATTACTGGGGCCAAGGCACCCTGGTGACTGTTAGCTCASEQ ID NO: 153DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYQWLTTFGQGTKVEIK SEQ ID NO: 154GATATCCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCCAGCGTGGGCGATCGCGTGACCATTACCTGCAGAGCCAGCCAGTCTATTTCTAACCGTCTGAACTGGTACCAGCAGAAACCGGGCAAAGCGCCGAAACTATTAATCTACAAAGGTTCTACTCTGCAAAGCGGCGTGCCGAGCCGCTTTAGCGGCAGCGGATCCGGCACCGATTTCACCCTGACCATTAGCTCTCTGCAACCGGAAGACTTTGCGACCTATTATTGCCAGCAGCATTACCAGTGGCTGACTACCTTTGGCCAGGGCACGAAAGTTGAAATTAAA SEQ ID NO: 155CAGGTGCAATTGGTGCAGAGCGGTGCCGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTTAGCTGCAAAGCATCCGGAGGGACGTTTTCTACTTTCTCTATCTCTTGGGTGCGCCAGGCCCCGGGCCAGGGCCTCGAGTGGATGGGCGGTATCATCCCGATCTTCGGCACTGCGAACTACGCCCAGAAATTTCAGGGCCGGGTGACCATTACCGCCGATGAAAGCACCAGCACCGCCTATATGGAACTGAGCAGCCTGCGCAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGGTGGTTACGGTGGTTACTACTACTTCGATTACTGGGGCCAAGGCACCCTGGTGACTGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAASEQ ID NO: 156DIQMTQSPSSLSASVGDRVTITCRASQSISNRLNWYQQKPGKAPKLLIYKGSTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYQWLTTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 157GATATCCAGATGACCCAGAGCCCGAGCAGCCTGAGCGCCAGCGTGGGCGATCGCGTGACCATTACCTGCAGAGCCAGCCAGTCTATTTCTAACCGTCTGAACTGGTACCAGCAGAAACCGGGCAAAGCGCCGAAACTATTAATCTACAAAGGTTCTACTCTGCAAAGCGGCGTGCCGAGCCGCTTTAGCGGCAGCGGATCCGGCACCGATTTCACCCTGACCATTAGCTCTCTGCAACCGGAAGACTTTGCGACCTATTATTGCCAGCAGCATTACCAGTGGCTGACTACCTTTGGCCAGGGCACGAAAGTTGAAATTAAACGTACGGTGGCCGCTCCCAGCGTGTTCATCTTCCCCCCCAGCGACGAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAAAGCGTCACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGT SEQ ID NO: 158QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGNIIPITGQTYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 159CAGGTGCAGCTGGTGCAGTCAGGCGCCGAAGTGAAGAAACCCGGCTCTAGCGTGAAAGTCAGCTGTAAAGCTAGTGGCGGCACCTTCTCTAGCTACGCTATTAGCTGGGTCAGACAGGCCCCAGGTCAGGGCCTGGAGTGGATGGGCAATATTATCCCTATCACCGGTCAGACCTACTACGCTCAGAAATTTCAGGGTAGAGTGACTATCACCGCCGACGAGTCTACTAGCACCGCCTATATGGAACTGTCTAGCCTGAGATCAGAGGACACCGCCGTCTACTACTGCGCTAGAGCCGCCTATCACCCCCTGGTGTTCGATAACTGGGGTCAGGGCACCCTGGTCACCGTGTCTAGCGCTAGCACTAAGGGCCCCTCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCCGGCGGCACAGCTGCTCTGGGCTGCCTGGTCAAGGACTACTTCCCTGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACCTCTGGCGTGCACACCTTCCCTGCCGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTGGTCACAGTGCCTTCAAGCAGCCTGGGCACCCAGACCTATATCTGCAACGTGAACCACAAGCCTTCCAACACCAAGGTGGACAAGCGGGTGGAGCCTAAGTCCTGCGACAAGACCCACACCTGTCCTCCCTGCCCTGCTCCTGAAGCTGCTGGCGGCCCTTCTGTGTTCCTGTTCCCTCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCTGAAGTGACCTGCGTGGTGGTGGACGTGTCCCACGAGGATCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAGGTGCACAACGCCAAGACCAAGCCTCGGGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTACAAGTGCAAAGTCTCCAACAAGGCCCTGCCTGCCCCTATCGAAAAGACAATCTCCAAGGCCAAGGGCCAGCCTAGGGAACCCCAGGTGTACACCCTGCCACCCAGCCGGGAGGAAATGACCAAGAACCAGGTGTCCCTGACCTGTCTGGTCAAGGGCTTCTACCCTTCCGATATCGCCGTGGAGTGGGAGTCTAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGCAAGSEQ ID NO: 160QSVLTQPPSASGTPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAISGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 161CAGTCAGTCCTGACTCAGCCCCCTAGCGCTAGTGGCACCCCTGGTCAGAGAGTGACTATTAGCTGTAGCGGCTCTAGCTCTAATATCGGTAATCACTACGTGAACTGGTATCAGCAGCTGCCCGGCACCGCCCCTAAGCTGCTGATCTATAGAAACAATCACCGGCCTAGCGGCGTGCCCGATAGGTTTAGCGGATCTAAGTCAGGGACTAGCGCTAGTCTGGCTATTAGCGGCCTGCAGTCAGAGGACGAGGCCGACTACTACTGTCAGTCCTGGGACTATAGCGGCTTTAGCACCGTGTTCGGCGGAGGCACTAAGCTGACCGTGCTGGGTCAGCCTAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAGCTGCAGGCCAACAAGGCCACCCTGGTGTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAAAAGACCGTGGCCCCAACCGAGTGCAGC SEQ ID NO: 162 GGSISTGSYSEQ ID NO: 163 QSPGY SEQ ID NO: 164 TTRYWMSHILAYGMDY SEQ ID NO: 165SSSNIGNHY SEQ ID NO: 166 ANT SEQ ID NO: 167 YDGSQSI SEQ ID NO: 168EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIWHSGPTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 169GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTTGGCATTCTGGTCCTACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA SEQ ID NO: 170DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVSWYQQLPGTAPKLLIYANTKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCSSYDGSQSIVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 171GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 172EVQLQESGPGLVKPGETLSLTCTVSGGSISTGSYYWNWIRQAPGKGLEWIGEIHGHGFTFYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARTTRYWMSHILAYGMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 173GAGGTGCAATTGCAAGAAAGTGGTCCGGGCCTGGTGAAACCGGGCGAAACCCTGAGCCTGACCTGCACCGTTTCCGGAGGTAGCATTTCTACTGGTTCTTATTATTGGAATTGGATTCGCCAGGCCCCTGGGAAGGGTCTCGAGTGGATTGGCGAGATTCATGGTCATGGTTTTACTTTTTATAATCCTTCTCTTAAGTCTCGGGTGACCATTAGCGTTGATACTTCGAAAAACCAGTTTAGCCTGAAACTGAGCAGCGTGACGGCGGCGGATACGGCCGTGTATTATTGCGCGCGTACTACTCGTTATTGGATGTCTCATATTCTTGCTTATGGTATGGATTATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA SEQ ID NO: 174DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVSWYQQLPGTAPKLLIYANTKRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCSSYDGSQSIVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 175GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGTCTTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATGCTAATACTAAGCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCTCTTCTTATGATGGTTCTCAGTCTATTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 176EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPHYGFAYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 177GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATTATTCCTCATTATGGTTTTGCTTATTATGCTCAGAAGTTTCAGGGTCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAASEQ ID NO: 178DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 179GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 180EVQLVQSGAEVKKPGSSVKVSCKASGGTFNSYAISWVRQAPGQGLEWMGNIIPYSGFAYYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARAAYHPLVFDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 181GAGGTGCAATTGGTTCAGTCTGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCCTCCGGAGGCACTTTTAATTCTTATGCTATTTCTTGGGTGCGCCAAGCCCCTGGGCAGGGTCTCGAGTGGATGGGCAATATTATTCCTTATTCTGGTTTTGCTTATTATGCTCAGAAGTTTCAGGGTCGGGTGACCATTACCGCGGATGAAAGCACCAGCACCGCGTATATGGAACTGAGCAGCCTGCGTAGCGAAGATACGGCCGTGTATTATTGCGCGCGTGCTGCTTATCATCCTCTTGTTTTTGATAATTGGGGCCAAGGCACCCTGGTGACGGTTAGCTCAGCCTCCACCAAGGGTCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCAGCGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGGGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAASEQ ID NO: 182DIVLTQPPSVSGAPGQRVTISCSGSSSNIGNHYVNWYQQLPGTAPKLLIYRNNHRPSGVPDRFSGSKSGTSASLAITGLQSEDEADYYCQSWDYSGFSTVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS SEQ ID NO: 183GATATCGTGCTGACCCAGCCGCCTTCAGTGAGTGGCGCACCAGGTCAGCGTGTGACCATCTCGTGTAGCGGCAGCAGCAGCAACATTGGTAATCATTATGTGAATTGGTACCAGCAGTTGCCCGGGACGGCGCCGAAACTTCTGATTTATCGTAATAATCATCGTCCCTCAGGCGTGCCGGATCGTTTTAGCGGATCCAAAAGCGGCACCAGCGCGAGCCTTGCGATTACGGGCCTGCAAAGCGAAGACGAAGCGGATTATTATTGCCAGTCTTGGGATTATTCTGGTTTTTCTACTGTGTTTGGCGGCGGCACGAAGTTAACCGTCCTAGGTCAGCCCAAGGCTGCCCCCTCGGTCACTCTGTTCCCGCCCTCCTCTGAGGAGCTTCAAGCCAACAAGGCCACACTGGTGTGTCTCATAAGTGACTTCTACCCGGGAGCCGTGACAGTGGCCTGGAAGGCAGATAGCAGCCCCGTCAAGGCGGGAGTGGAGACCACCACACCCTCCAAACAAAGCAACAACAAGTACGCGGCCAGCAGCTATCTGAGCCTGACGCCTGAGCAGTGGAAGTCCCACAGAAGCTACAGCTGCCAGGTCACGCATGAAGGGAGCACCGTGGAGAAGACAGTGGCCCCTACAGAATGTTCA SEQ ID NO: 184 GGSISTGSYSEQ ID NO: 185 NHMGI SEQ ID NO: 186 TTRYWMSHILAYGMDY SEQ ID NO: 187SSSNIGNHY SEQ ID NO: 188 ANT SEQ ID NO: 189 YDGSQSI SEQ ID NO: 190GGSISTGSY SEQ ID NO: 191 WHSGP SEQ ID NO: 192 TTRYWMSHILAYGMDYSEQ ID NO: 193 SSSNIGNHY SEQ ID NO: 194 ANT SEQ ID NO: 195 YDGSQSISEQ ID NO: 196 EIHGHGFTFYNPSLKS SEQ ID NO: 197 GGSISTGSY SEQ ID NO: 198HGHGF SEQ ID NO: 199 TTRYWMSHILAYGMDY SEQ ID NO: 200 SSSNIGNHYSEQ ID NO: 201 ANT SEQ ID NO: 202 YDGSQSI SEQ ID NO: 203 GGTFNSYSEQ ID NO: 204 IPIYGT SEQ ID NO: 205 AAYHPLVFDN SEQ ID NO: 206 SSSNIGNHYSEQ ID NO: 207 RNN SEQ ID NO: 208 WDYSGFST SEQ ID NO: 209 GGTFNSYSEQ ID NO: 210 NPFYIGE SEQ ID NO: 211 AAYHPLVFDN SEQ ID NO: 212SSSNIGNHY SEQ ID NO: 213 RNN SEQ ID NO: 214 WDYSGFST SEQ ID NO: 215GGTFNSY SEQ ID NO: 216 IPHYGF SEQ ID NO: 217 AAYHPLVFDN SEQ ID NO: 218SSSNIGNHY SEQ ID NO: 219 RNN SEQ ID NO: 220 WDYSGFST SEQ ID NO: 221GGTFNSY SEQ ID NO: 222 IPYSGF SEQ ID NO: 223 AAYHPLVFDN SEQ ID NO: 224SSSNIGNHY SEQ ID NO: 225 RNN SEQ ID NO: 226 WDYSGFST SEQ ID NO: 227GGTFNSY SEQ ID NO: 228 NPFYIGE SEQ ID NO: 229 AAYHPLVFDN SEQ ID NO: 230SSSNIGNHY SEQ ID NO: 231 RNN SEQ ID NO: 232 WDYSGFST SEQ ID NO: 233GGTFNSY SEQ ID NO: 234 IPMTGQ SEQ ID NO: 235 AAYHPLVFDN SEQ ID NO: 236SSSNIGNHY SEQ ID NO: 237 RNN SEQ ID NO: 238 WDYSGFST SEQ ID NO: 239NIIPITGQTYYAQKFQG SEQ ID NO: 240 GGTFSTF SEQ ID NO: 241 IPIFGTSEQ ID NO: 242 GGYGGYYYFDY SEQ ID NO: 243 SQSISNR SEQ ID NO: 244 KGSSEQ ID NO: 245 HKVWLT SEQ ID NO: 246 GGTFSTF SEQ ID NO: 247 IPIFGTSEQ ID NO: 248 GGYGGYYYFDY SEQ ID NO: 249 SQSISNR SEQ ID NO: 250 KGSSEQ ID NO: 251 HYVWST SEQ ID NO: 252 GGTFSTF SEQ ID NO: 253 IPIFGTSEQ ID NO: 254 GGYGGYYYFDY SEQ ID NO: 255 SQSISNR SEQ ID NO: 256 KGSSEQ ID NO: 257 HYQWLT SEQ ID NO: 258 GFTFSSY SEQ ID NO: 259 QSSGENSEQ ID NO: 260 VMIGYGFDY SEQ ID NO: 261 SQSIFNY SEQ ID NO: 262 DSSSEQ ID NO: 263 YSGLLF

1-48. (canceled)
 49. A method of treating and/or preventing sarcoidosis,hemophagocytic lymphohistiocytosis (HLH), familial hemophagocyticlymphohistiocytosis (FHL) and other immunodeficiency syndromes, GiantCell Arthritis (GCA), chronic obstructive pulmonary disease (COPD),adult onset Still's Disease (AOSD), systemic juvenile idiopathicarthritis (SJIA), severe asthma, Uveitis, Geographic Atrophy, diabetestype 1, diabetes type 2 or atherosclerosis and any combination thereofin a mammalian patient which method comprises administrating to themammalian patient a therapeutically effective amount of an isolatedantibody or fragment thereof that specifically binds IL-18 comprising:i. a heavy chain variable region H-CDR1 comprising SEQ ID NO: 3 and ii.a heavy chain variable region H-CDR2 comprising SEQ ID NO: 4 or SEQ IDNO: 9 or SEQ ID NO: 10 or SEQ ID NO: 11 or SEQ ID NO: 12 or SEQ ID NO:13 and iii. a heavy chain variable region H-CDR3 comprising SEQ ID NO: 5and iv. a light chain variable region L-CDR1 comprising SEQ ID NO: 6 andv. a light chain variable region L-CDR2 comprising SEQ ID NO: 7 and vi.a light chain variable region L-CDR3 comprising SEQ ID NO:
 8. 50. Amethod of treating and/or preventing sarcoidosis, hemophagocyticlymphohistiocytosis (HLH), familial hemophagocytic lymphohistiocytosis(FHL) and other immunodeficiency syndromes, Giant Cell Arthritis (GCA),chronic obstructive pulmonary disease (COPD), adult onset Still'sDisease (AOSD), systemic juvenile idiopathic arthritis (SJIA), severeasthma, Uveitis, Geographic Atrophy, diabetes type 1, diabetes type 2 oratherosclerosis and any combination thereof in a mammalian patient whichmethod comprises administrating to the mammalian patient atherapeutically effective amount of a pharmaceutical compositioncomprising an isolated antibody or fragment thereof that specificallybinds IL-18 comprising: i. a heavy chain variable region H-CDR1comprising SEQ ID NO: 3 and ii. a heavy chain variable region H-CDR2comprising SEQ ID NO: 4 or SEQ ID NO: 9 or SEQ ID NO: 10 or SEQ ID NO:11 or SEQ ID NO: 12 or SEQ ID NO: 13 and iii. a heavy chain variableregion H-CDR3 comprising SEQ ID NO: 5 and iv. a light chain variableregion L-CDR1 comprising SEQ ID NO: 6 and v. a light chain variableregion L-CDR2 comprising SEQ ID NO: 7 and vi. a light chain variableregion L-CDR3 comprising SEQ ID NO:
 8. 51. The method according to claim49, wherein the mammalian patient is a human patient.
 52. The methodaccording to claim 50, wherein the mammalian patient is a human patient.53. A method for detecting and/or measuring the presence and/or amountof free IL-18 (i.e. IL-18 not bound to IL-18BP) in a sample, wherein thesample is optionally a human sample, wherein the method comprisescontacting the sample with an isolated antibody or a fragment thereofthat specifically binds IL-18 comprising: i. a heavy chain variableregion H-CDR1 comprising SEQ ID NO: 3 and ii. a heavy chain variableregion H-CDR2 comprising SEQ ID NO: 4 or SEQ ID NO: 9 or SEQ ID NO: 10or SEQ ID NO: 11 or SEQ ID NO: 12 or SEQ ID NO: 13 and iii. a heavychain variable region H-CDR3 comprising SEQ ID NO: 5 and iv. a lightchain variable region L-CDR1 comprising SEQ ID NO: 6 and v. a lightchain variable region L-CDR2 comprising SEQ ID NO: 7 and vi. a lightchain variable region L-CDR3 comprising SEQ ID NO:
 8. 54. The methodaccording to claim 53, wherein the sample is blood, optionally humanblood.
 55. A diagnostic kit comprising an isolated antibody or fragmentthereof that specifically binds IL-18 comprising: i. a heavy chainvariable region H-CDR1 comprising SEQ ID NO: 3 and ii. a heavy chainvariable region H-CDR2 comprising SEQ ID NO: 4 or SEQ ID NO: 9 or SEQ IDNO: 10 or SEQ ID NO: 11 or SEQ ID NO: 12 or SEQ ID NO: 13 and iii. aheavy chain variable region H-CDR3 comprising SEQ ID NO: 5 and iv. alight chain variable region L-CDR1 comprising SEQ ID NO: 6 and v. alight chain variable region L-CDR2 comprising SEQ ID NO: 7 and vi. alight chain variable region L-CDR3 comprising SEQ ID NO: 8, wherein thekit optionally comprises a first control compound.
 56. The diagnostickit according to claim 55 wherein the first control compound is freeIL-18 and wherein the kit optionally comprises a second control compoundwherein the second control compound is murine antibody 125-2H.
 57. Amedical or diagnostic device comprising an isolated antibody or afragment thereof that specifically binds IL-18 comprising: i. a heavychain variable region H-CDR1 comprising SEQ ID NO: 3 and ii. a heavychain variable region H-CDR2 comprising SEQ ID NO: 4 or SEQ ID NO: 9 orSEQ ID NO: 10 or SEQ ID NO: 11 or SEQ ID NO: 12 or SEQ ID NO: 13 andiii. a heavy chain variable region H-CDR3 comprising SEQ ID NO: 5 andiv. a light chain variable region L-CDR1 comprising SEQ ID NO: 6 and v.a light chain variable region L-CDR2 comprising SEQ ID NO: 7 and vi. alight chain variable region L-CDR3 comprising SEQ ID NO:
 8. 58. Themethod of claim 49 wherein the method comprises treating and/orpreventing pulmonary sarcoidosis.
 59. The method of claim 50 wherein themethod comprises treating and/or preventing pulmonary sarcoidosis. 60.The method of claim 49, wherein the antibody or fragment thereofcomprises a heavy chain variable region H-CDR2 comprising SEQ ID NO: 9.61. The method of claim 49, wherein the antibody or fragment thereofcomprises a heavy chain variable region H-CDR2 comprising SEQ ID NO: 13.62. The method of claim 49, wherein the isolated antibody or fragmentcomprises a light chain variable domain comprising SEQ ID NO: 16 or SEQID NO:
 20. 63. The method of claim 49, wherein the isolated antibody orfragment thereof comprises: i. a heavy chain variable domain comprisingSEQ ID NO: 14 or SEQ ID NO: 22 or SEQ ID NO: 25 or SEQ ID NO: 28 or SEQID NO: 31 or SEQ ID NO: 34 and a light chain variable domain comprisingSEQ ID NO: 16 or conservative variants thereof or ii. a heavy chainvariable domain comprising SEQ ID NO: 18 or SEQ ID NO: 37 or SEQ ID NO:40 and a light chain variable domain comprising SEQ ID NO:
 20. 64. Themethod of claim 49, wherein the isolated antibody or fragment thereofcomprises a heavy chain variable domain comprising SEQ ID NO: 14 and alight chain variable domain comprising SEQ ID NO:
 16. 65. The method ofclaim 49, wherein the isolated antibody or fragment thereof amino acidlysine (Lys; K) in position 30 with reference to SEQ ID NO:14 isreplaced by an amino acid selected from asparagine (Asn; N) or serine(Ser; S) or threonine (Thr; T) or alanine (Ala; A) or glutamate (Glu; E)or histidine (His; H) or leucine (Leu; L) or glutamine (Gln; Q) orarginine (Arg; R) or valine (Val; V) or tyrosine (Tyr; Y) or isoleucine(Ile; I).
 66. The method of claim 63, wherein the isolated antibody orfragment thereof comprises a heavy chain variable domain comprising SEQID NO: 18 and a light chain variable domain comprising SEQ ID NO: 20.67. The method of claim 49, wherein the isolated antibody comprises: i.a heavy chain comprising SEQ ID NO: 43 or SEQ ID NO: 47 or SEQ ID NO: 50or SEQ ID NO: 56 and a light chain comprising of SEQ ID NO: 45 or ii. aheavy chain comprising SEQ ID NO: 53 or SEQ ID NO: 100 or SEQ ID NO: 158thereof and a light chain comprising SEQ ID NO:
 160. 68. The method ofclaim 67, wherein the isolated antibody comprises: i. a heavy chaincomprising SEQ ID NO: 43 thereof and a light chain comprising of SEQ IDNO: 45 or ii. a heavy chain comprising SEQ ID NO: 158 and a light chaincomprising SEQ ID NO: 160.